Table of content

Jornal of Biotechnology Research Center

مجلة مركز بحوث التقنيات الاحيائية

ISSN: 18151140
Publisher: Al-Nahrain University
Faculty: Presidency of the university or centers
Language: Arabic and English

This journal is Open Access

About

The Biotechnology Research Center (BRC) in a Al-Nahrain University issued in 2007 the first edition of a tightly seasonal scientific journal named as the journal Biotechnology Research Center (JBRC) which got authorization in 2005 and held an impact number ISSN: 1815-1140.The journal accepts scientific researches in Arabic and English.
JBRC’s main interest is Biotechnology researches in the Medical, Molecular, Agriculture and environmental fields which have an important impact on the public and private sectors in Iraq.
JBRC’s structure consists of editing committee (headed by the manger of the BRC and the editor in chief), consulting committee (contains a well- known iraqi scientists in biotechnology) and editing secretary.
All researches are submitted to the JBRC’s regulations which is mainly is that they must be according to the journal directions and instructions , then the researches will be evaluated by three well-known scientists in the field and after that reviewing occurred by the editing committee to ensure and verify all JBRC’s instructions and regulations are taken into consideration .
A special edition of JBCR is issued to cover all researches that presented to the BRC’s scientific conferences which subjected to all regulations and instructions of publishing in JBCR.

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brcn2012@yahoo.comيتم الاتصال عبر البريد الالكتروني لمجلة مركز بحوث التقنيات الاحيائية


0096407707766148او للاتصال تلفونيا
بسكرتارية التحرير
م.م. سعاد محمد مجيدب
محمد منير حسين

Table of content: 2011 volume:5 issue:1

Article
Identification strains of Bean Yellow Virus potyvirus (BYMV) on bean Phaseolus vulgaris in Iraq
تشخيص سلالات فايروس موزائيك الفاصوليا الأصفر (BYMV)Bean yellow mosaic virus potyvirusعلى نباتات الفاصوليا Phaseolus vulgarisفي العراق

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Abstract

everal methods were used to identify the virus, such as indicator plants, physical properties, manners transmission by aphids and ELISA test and among indicator plants two strains of BYMV were identified. The first one was showed a necrotic local lesion on bean while the second one was showed strong epinasty, veinal necrosis in inoculated leaves then it caused top necrosis at last severe systemic mottle. This two different symptoms gives a positive reaction at ELISA test and this test was indicated that the two symptoms is belong for two strains of bean yellow mosaic virus ( BYMV).The physical properties for the first strain about the Thermal Inactivating point (TIP) was 50-55ْc while the second strain was 60ْc and the dilution end point (DEP)for the first strain was 10-2 but it was 10-2 -10-3 for the second one and the longevity in vitro (LIV) for the first strain 48 hrs at room temperature and the main transmission by insect was achieved by several species of aphids.The double immunodiffution test in agar have been showed meeting of two precipitin line without cross in this two lines when used antiserum of the first strain of virus against the two sap of infected plants with both strains of virus and this test was used in the second strain against sap of the both infected plants too with the two strains of the virus and this test was additional indication that the two symptoms in bean were belonged to a two different strains to the same virus.

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Article
Cytotoxic effect of the aqueous extract of Portulaca Oleracea L. on some cell lines
السميه الخلوية للمستخلص المائي لنبات البقلة Portulaca Oleracea L. لبعض الخطوط الخلوية

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Abstract

he cytotoxic effect of the aqueous extract of Portulaca oleracea L. was tested in vitro against the cancer cell AMN3 (Murine Mammary Adenocarcinoma cell line) and the normal cell REF (Rat Embryo Fibroblasts). Proliferation was significantly reduced at high concentrations (300-1000 µg/ml) during the first 24 hrs. of contact on AMN3. while the extract showed no significant effect during different period time of exposure as compared with control against REF cells.

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Article
Effects of propolis extract on growth of Entamoeba histolytica (trophozoites) in vitro
تأثير المستخلص الكحولي للعكبر على نمو الأطوار المتغذية للأميبا الحالة للنسج في الزجاج

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Abstract

Amoebiasis is the third leading cause of death from parasitic disease worldwide. The causative protozoan parasite, Entamoeba histolytica, is a potent pathogen. Propolis is a natural resinous substance collected by bees from vegetable sources and it has a therapeutic properties have been investigated in this work. So we evaluated the inhibitory activity of ethanolic extracts of propolis (EEP) on Entamoeba histolytica trophozoite growth. Propolis inhibited the growth of E.histolytica trophozoites and the level of inhibition varied according to the extract concentration and incubation times, it also showed a marked activity on cell lysis of trophozoite. The highest reduction of parasite growth was observed in culture exposed to (25,50 mg/ml) of propolis, in all incubation periods (24,48 )h. Growth reduction by 76 % was observed in 12.5 mg/ml propolis-treated culture, while the concentrations of (25, 50) mg/ml were able to inhibit growth by more than 90%. Light microscopic study showed morphological changes in E.histolytica trophozoites.


Article
Effect of methanolic extract for (leaves and roots) of Bacopa monniera L. aerial plant parts on the growth of some bacteria and fungi
تأثير المستخلص الكحولي للأجزاء الهوائية (الاوراق والسيقان) لنبات البربين البري Bacopa monniera في نمو بعض انواع البكتيريا والفطريات

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Abstract

he project was conducted to study the inhibitory activity of methanolic extract for aerial parts (leaves and roots) of Bacopa monniera L. on some bacterial species represented by: Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Escherchie coli, Klebsellia sp. Shigella sp., by using the Agar-well diffusion method for estimating the Minimum Inhibition Concentration (MIC) and Minimum Bactericidal Concentration (MBC) as well as two type of fungi, Aspergillus niger and Aspergillus flavus, the fungi used to study the antifungal activity of plant extract. The results showed a difference in the effect for the types of bacteria as the value of (MIC) for the types Bacillus cereus, Shigella sp. and Klebsellia sp. Is a (100, 200, 200) while there was no effect of the extract against other bacterial species under study. As for the fungi testing, the extract has a clear effect by measuring the mean of growth diameter for fungi, where the growth diameter of the fungus A. niger 1.03 cm at the concentration 50mg/ml while the diameter became 0.5 cm at the 300 mg/ml concentrate in the first day, and in the seventh day of the experiment, the growth diameter reached 7.03 cm at the 50 mg/ml concentrate but the concentration 300mg/ml were 6.4 cm compared with control, which amounted to 7.6 cm. As for the fungi A. flavus growth diameter (1.1 , 0.46) cm at concentrations (50, 300) mg/ml in the first day of the experiment, in the seventh day the diameter was (4.73,3.9) cm at concentrations (50, 300) mg/ml respectively, while the diameter of growth has been different at the other concentrations and within test days.

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Article
Detection of toxigenic Vibrio cholerae by PCR
الكشف عن بكتريا ضمات الكوليرا المنتجة للسموم باستخدام الـPCR

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Abstract

holera toxin (CT) is a major virulence factor of V. cholerae causing water diarrhea. The detection of CT-producing V. cholerae using conventional culture-, biochemical- and immunological-based assays is time-consuming, laborious, and requiring more than three days perform. In this work a specific primers for ctxB gene were used for detection of V. cholera in water samples. Few colonies of V. cholera were suspended in water and used as a template in PCR reaction for the detection of ctxB gene. The 391-bp sequence of a gene that codes for the cholera toxin B subunit was amplified by PCR. Direct use of V. cholerae pure culture for PCR replaces the need for DNA extraction or boiling. Increase the concentration of MgCl2 enhances the efficiency of amplification. The specificity of the assay was determined to be specific for V. cholerae but not for, vibrio related bacteria, E. coli, Non-Agglutinable (NAG) V. cholerae, and Aeromonas sp.

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Article
Using of Data Base to Determine the Restriction Sites and Drawing Restriction Map for Lipase Gene from Bacillus stearothermophilus
استخدام قواعد البيانات لتحديد مواضع القطع ورسم خارطة التقييد للجين المشفر لأنزيم اللايبيز من بكتريا Bacillus stearothermophilus

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Abstract

pecific data base was used for restriction enzymes (rebase) and related proteins, to design executive program in quick basic language to determine the restriction sites and drawing restriction map for the complete sequence of lipase gene from Bacillus stearothermophilus. This program which was of a great benefit in prediction of the restriction patterns for the analysis of recombinant DNA molecules was used to determine the restriction sites for 16 enzyme among 46 type of different restriction enzymes having recognition sequences included in lipase gene which they are SmaI, XhoI, SalI, DdeI, ClaI, EagI, BspHI, AccI, EcoRV, HaeII, BanI, BclI, CfrI , SstI, KspI and NaeI. Results showed that it could be determine accurately the restriction sites and fragments sizes resulted in case of the treatment of lipase gene with these restriction endonucleases. It was also enabled to draw restriction map specific for these enzymes and determine coding region for lipase gene sized of 1254 bp included in the complete sequence for the cDNA of the gene sized of 1725 bp.

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Article
Effect of Iraqi probiotic supplementation on body Weight Gain and some blood parmeters of Awassi Lambs fed different
تأثير اضافة المعزز الحيوي العراقي الى علائق الحملان العواسية المعرضة لنسب مختلفة من سايلج القصب و دريس الجت في بعض متغيرات الدم

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Abstract

in this study, twenty four individual Awassi male lambs (mean body weight 17.05 kg and 3-4 months of old) were used to investigate the effect of three ratios of alfalfa hay to reed silage (40:0, 20: 20 and 0: 40 AH: RS ratios) fed with two levels of Iraqi probiotic (IP) (0 and 7.5 g IP / kg DM) on blood cholesterol concentration (BCC), blood urea nitrogen concentration (BUNC) and blood globulin concentration(BGC). The diets were formulated as DM to give a 40 parts alfalfa hay or/and reed silage and 60 parts concentrate. Within 2-3 days before end of the period trial, blood samples were taken during 0, 6, 12 and 24 hr. post morning feeding for measuring BCC, BUNC and BGC. No differences among treatments were found in daily feed intake for all nutrients. Different AH: RS ratios have no effect on live weight gain (LWG), BCC, BUNC and BGC. However, lambs fed diets supplemented with IP tended to increase in their LWG and significantly reduce BGC and BUNC (P<0.05 and P<0.01 respectively) as compared with those fed diets without IP. While, IP supplementation had no effect on BCC. In conclusion, during 0, 6, 12 and 24 hr. post morning feeding AH: RS ratios had no effect on BCC, BUNC and BGC. Inconclusion, IP supplementation was associated with higher LWG and lower BUN concentration.


Article
The effect of boron foliar on the germination and vigor of the resultant seed of sunflower (Helianthus annuus L.) plants
تأثير رش نباتات زهرة الشمس (Helianthus annuus L.) بالبورون في انبات وقوة البذور الناتجة منها

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Abstract

Laboratory study was carried out on sunflower seed products resulting from the sunflower plants foliar by different concentrations of boron (0, 50, 100, 150, 200, 250) ppm at flowering stage in spring and autumn seasons in 2009 at the laboratories of the Ministry of Agriculture - the Authority for public inspection and certification seeds –Karbala branch to study the effect of post-boron foliar on the germination of seeds, radical length, plumule length and dry weight of seedling. An analysis of variance according to complete randomized design with four replicates were used. Simple correlation between the attributes were examined. a regression analysis was performed, and prediction equation of the attributes was studied. The results showed that seeds from plants sprayed with boron concentration of 150 ppm was superior in all characters, the first count, final count, radical length, plumule length, dry weight of seedling and cold test for both seasons (82.25% and 92.00% and 122.75 mm and 89.00 mm and 61.00 mg and 88.00 %) respectively, in the spring season, and (76.25% and 80.00% and 118.75 mm and 100.50 mm and 68.00 mg and 72.00%) respectively in the autumnal season. Results showed significant positive correlation between most of the studied traits. Regression analysis showed the possibility of calculating the expected equation of equivalent boron which is significant in the proportion of normal seedling in standard germination test and plumule length in spring season, and the proportion of normal seedling in standard germination test, radical length and a proportion of normal seedling in cold test in autumn season.

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Article
Production of Ethanol from Sugars Fermentation By Yeasts Using Bioreactor
استخدام الخمائر في انتاج الايثانول من تخمر السكر وباستعمال المفاعل الحيوي

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Two yeast isolates of C.kefyr-T and C.kefyr-S, were isolated from crude whey, the optimum pH value was 5, and S.cerevisiae was significantly best in ethanol and Biomass production from all glucose concentrations compared to C.kefyr-T and C.kefyr-S which were significantly best ethanol and biomass production from all galactose concentrations comparing with S.cerevisiae. C.kefyr-T which was best in production compared to C.kefyr-S. Best ethanol production was 8.7% Ethanol from 10% sugar mixture, by mixed culture of (S. cerevisiae+ C.Kefyr-T), and best ethanol production was 4.6% from 8% synthetic lactose by mixed culture of (C.kefyr-T+C.kefyr-S). There is a significant decrease in ethanol and biomass production from all mixtures by using mixed culture of (C.kefyr-T+ C.kefyr-S+ S. cerevisiae) than using mixed culture of (C. kefyr + S. cerevisiae) and mixed culture of (C.kefyr-T + C.kefyr-S). Different period of times at (24, 36, 48, and 60) hof fermentation in bioreactor are presented. The initial sugar mixture concentration was 10% in all cases. The ethanol production during the 48 h gave a maximum value of 15.5 % by mixed culture of (C. kefyr + S. cerevisiae), but it remains at low levels within other periods, whereas the highest biomass16% gained at the same bioreactor conditions.

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Article
Characterization of purified Polyphenol oxidase (PPO) from (Lactuca serriola L.) Prickly lettuce
توصيف أنزيم (PPO)Polyphenol oxidase المنقى من نبات الخس الشوكي Prickly lettuce (Lactuca serriola L.)

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Abstract

n this study, purified PPO from Prickly lettuce were obratind from a previous characterization study. The results showed that the optimum pH were 6.5 and the pH stability profile showed that the enzyme were more stable at pH values 5.5-8, the optimum temperature for enzyme activity were 35ºC and the enzyme were stable at 35ºC for 10 min. the activation energy for conversion of substrate to product was 39.7194 Kj/mol, the purified enzyme lost its activity after 5 days of storage at 25ºC, while it lost (73 , 16)% of primary activity after 21 days of storage at 4 and -18ºC respectively, the purified enzyme contains 7.2% carbohydrates. It was noticed that all treated of enzyme with L-Cystein, Sodium metabisulphate and Dithiothretol at 1 and 10 mM revealed high inhibition for the enzyme activity, followed by citric acid, L-Ascorbic acid, EDTA and Potassium cyanide, while both Thiourea and Sodium benzoate showed less inhibitory effect. The Km and Vmax were (4.762 , 0.0588) mM/min respectively, while the coefficient Vmax/Km were 0.01235 min-1 for the purified enzyme when used Catechol as a substrate.

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Article
Bacteriophages effects on antibiotic sensitivity of Staphylococcus aureus
تأثير العاثيات على حساسية بكتريا المكورات العنقودية الذهبية تجاه مضادات الحيوية

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A total of 50 swab samples were collected from patients suffering from different infections. Morphological and biochemical characterization showed that 46% isolates were characterized as Staphylococcus aureus. All these isolates were able to grow on mannitol salt agar medium and ferment mannitol and sucrose. β- hemolysin, coagulase production and antibiotics sensitivity of S.aureus, before and after treatment with phage, was examined. It showed there is no change in the β- hemolysin and in the coagulase production but lysogenic phage effect the antibiotic sensitivity of S. aureusisolates significantly. In addition plasmid DNA was isolated from bacteria whose showed multiple antibiotic resistances. Large and small bands were obtained .Thereafter curing process had done to evaluate the plasmids content, it was found entire losing for the small plasmids, while the large plasmids remain intact. From this study, it can be concluded that S.aureusisolates found in wound at high percentage; moreover,S.aureus treated with phages effect the sensitivity patterns against antibiotics.


Article
Production an immuno kit for identification two strains of Ralstonia solancearum (smith) by ELISA which caused a bacterial wilt on tomato in Iraq
انتاج عدة تشخيصية مناعية للكشف عن سلالتين للبكتيريا Ralstonia solancearum (smith) بطريقة اليزا ELISA المسببة لمرض الذبول البكتيري على الطماطة في العراق

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R alstonia solancearum is one of most important tomato pathogen which annually causes considerable yield loss. Strains of these pathogenic bacteria were identified based on both phenotypic and differentiation media on Tetrazolium Chloride Agar (TZC) and the hypersensitivity tests on leaves of tobacco cultivar xanthi. Many biochemical adapted tests were carried out for identification of both the genus and species of the bacteria. Two strains R1 and R3 were identified. Antiserum of each strain was obtained following the injection of pure cell suspension of each strain in lab. Rabbit. Immunoglobulin (IgG) was purified and conjugated with alkaline phosphates and used as an identification kit for each strain by ELISA test .The reading of ELISA reader at 405 nanometer when testing plant extract infected by R1 strain with its antiserum were 1.14 while it was 1.10 for R3 strain, ELISA test were also carried out for plant extract infected by R1 strain and the antiserum of R3 strain, ELISA test were carried out too for plant infected by R3 strain and antiserum of R1 strain the reading of ELISA reader were 0.90, 0.92 respectively. The result of ELISA test was negative when using the normal serum and carbonate buffer as antiserum. The result of this test is clear evidence showed the high specify of the antiserum which produced in this study.

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Article
Serum alpha-1-antitrypsin level in Iraqi patients with coronary heart disease
مستوى بروتين الفا-1-مضاد التربسين في مصل الدم عند مرضى عراقيين مصابين بامراض شرايين القلب الاكليلية

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T his study was designed to estimate the level of serum alpha -1- antitrypsin in patients with coronary heart disease. Patients were clinically subdivided into stable angina 25 patients, unstable angina30 patients and myocardial infarction 50 patients. A control sample of 30 individuals was matched with patient for age and sex.Non-significant elevation of serum alpha one antitrypsin were noted in MI cases compared to healthy control individuals, even some MI patients showed decreasing level of alpha one antitrypsin, which may return to hereditary alpha one antitrypsin deficiency in Iraqi patients.The Stable angina patients group showed non-significant decrease in alpha one antitrypsin level compared to healthy control, while unstable angina patients group showed significant decrease in alpha one antitrypsin level that will may facilitate developing disease towards MI. All present results need additional studies to be carried out on larger samples of Iraqi individuals and alpha one antitrypsin deficient patients, furthermore, highlighting on the relationship between alpha one antitrypsin and risk of coronary heart disease.


Article
The side effects (cytogenetics ) of Methotrexate drug on male germ cells of Mus musculus
التاثيرات الجانبية (الوراثية الخلوية) للعقار ميثوتريكسات Methotrexate في الخلايا الجنسية الذكرية للفأر المختبري Mus musculus

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I n this study three doses (0.01, 0.02, 0.04) mgkg B.W of methotrexate drug which used, in the treatment of bladder cancer, were investigate in male germ cells of Mus musculus. Five tests were considered (mitotic index, chromosomal central associated, micronuclei, chromosomal aberration and sperm–head abnormalities). The results showed that,s these doses were presentel with high inhibitory effect of cell division, as well as, they induced significantly higher number of chromosomal central associated, micronuclei, chromosomal aberration and sperm head abnormalities especially after (21, 35) days after treatment at equal rate in control. We study the relationship between the structures of drug with it medical effect by study the atomic charge of each atom of drug and adenine, guanine, cytosine and uracil.


Article
Identification of differences in virulence factors production from mutant isolates of clinical Vibrio cholerae S
تشخيص الاختلافات في انتاج عوامل الضراوة من عزلات طافرة للبكتريا المرضية Vibrio cholerae S

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A ntibiotic resistant mutants for rifampicin, streptomycin and klindamycin were isolated from the clinical isolate of Vibrio choleraeS mutated by chemical mutagens. Mutation frequency of V. cholerae S depends on the treatment time and the highest viable count of antibiotic resistant were for Rifampicin after treatment with Acridine orange, Ethedium bromide, Nitrosoguanidine, 5-Florouracil, 2-Bromouracil and cyclophosphamide. One thousand mutant isolates were examined for morphological differences in colony surface, color and diameter. The treatment with AO, NTG, 5-FU, and 2-BU gave opaque to orange color larger diameter about 5-7 mm of Rifampicin resistant mutant isolates at TSA and 25% of these mutant appeared as wrinkled surface. Klindamycin resistant mutants of V. cholerae S were appeared as similar to the wild type while, Streptomycin resistant mutants of appeared as pin- point white smooth colonies on TSA. No differences were seen for oxidase, string test and fermentation pattern for sucrose and lactose. Toxin CoregulatedPili production was differed from high level order designated as +++ to mild ++ and low level designated as + after mutation with 5-FU and 2-BU. However, 15% of rifampicin resistant mutant isolates gave no agglutination phenomena. No proteases activity detected even after 48 hour of incubation; the production of lipases enzyme did not affected; while, mutator isolates produced high level of β- haemolysins about 2.5 fold. About 90% of cyclophosphamide- rifampicin mutant showed homogenized culture with no auto agglutination but high level of proteases. While, only 10% of cyclophosphamide - rifampicin mutants gave slightly auto agglutination and didn’t produce proteases enzyme. The production of both TCP and CT were increased from rough pigment producing mutant isolates comparing with yellow and smooth mutants.

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Article
Molecular Diagnosis of Chronic Myeloid Leukemia & Monitoring Response to Different Types of Treatment
التشخيص الجزيئي لابيضاض الدم النقياني المزمن بواسطة تفاعل البلمرة الكمي المتسلسل و تقييم الاستجابة للعلاجات المختلفة

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C hronic myeloid leukemia (CML) is one of leukemia types which account for 15 % to 20% of all leukemia cases. Patients are presented with splenomegaly, fever, anemia, fatigue, weight loss, and weakness. It is results from reciprocal translocation (9; 22).This abnormality is called Philadelphia (Ph) chromosome and it is detected in 95% of patients with CML, and in 20% of patients with Acute Lymphocytic Leukemia (ALL). "Imatinibmesylate” is the most widely used drug for CML treatment because it targets the abnormal fusion gene. Blood samples were collected from (39) CML patients (19 males and 20 females) from July - November 2009 in The NationalCenter of Hematology/Baghdad. The age range were(8 – 70) years. According to the real time PCR results; the patients were divided into four groups: 1) PCR negative group. 2) PCR positive group on Hydroxyurea treatment 3) PCR positive group on Gleevec® treatment 4) PCR positive group with no treatment (recently diagnosed).Patients were selected randomly. Their RNA was extracted from peripheral blood cells and reverse transcribed into cDNA which was amplified using real time PCR to measure the ratio of BCR-ABL fusion gene in their Philadelphia chromosome. This is to confirm the diagnosis and evaluate the response to the most widely used drugs for CML treatment (Gleevec® 400 mg/d and hydroxyurea 450 mg/d for two months at least). This study excluded CML diagnosis in 10 patients, so other myeloproliferative disorders need to be verified. The group treated with Gleevec® showed a better response than hydroxyurea at the molecular level.Key words: leukemia, chronic myeloid leukemia, polymerase chain reaction, real time PCR, BCR-ABL, quantitative PCR, hydroxyurea, imatinibmesylate, Philadelphia chromosome, tyrosine kinase.

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