National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001Enzyme Immobilization: The State of Art in Biotechnology1972066902ENDariush NorouzianPilot Biotechnology Department, Pasteur Institute of Iran,Tehran, I.R. Iran.Journal Article20031001The advantages of immobilized enzyme over its soluble counterpart arise from their improved stability and<br />easy separation from the reaction media, leading to decrease in production cost. Immobilization methods<br />range from adsorption onto matrices, entrapment, cross-linking and covalent bonding to prefabricated<br />carriers or activated supports. Changes in kinetic properties of immobilized enzyme can produce substrate<br />or pH gradient, which reduce the reaction rates and finally product yields.<br /><br />National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001Purification of Large Quantities of Biologically Active Recombinant Human Growth Hormone2072126884ENMahvash KhodabandehNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Bagher YakhchaliNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.0000-0003-4031-877xMaryam RahimiNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Neda VaseliNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Zahra Moghaddasi JahromiNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Alireza ZomorrodipourNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.0000-0003-0671-4779Abdolkhalegh DeezagiNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Saeid Ansari MajdNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Mohammad H SanatiNational Research Center for Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran.Journal Article20031001Production and purification of human growth hormone using a simple method was studied in two recombinant<br />Escherichia coli, D7-5 and C27-2 strains. The r-hGH was expressed in the form of inclusion body in a batch<br />fermentation process and purified to 99% purity using a procedure based on acid precipitation of the host<br />derived proteins and other impurities. The effect of the pH and host strain on purification of the r-hGH and<br />efficiency of the procedure were evaluated. It was found that the optimum pH for precipitation of the host<br />derived proteins was 4.9. The procedure was suitable for r-hGH purification from D7-5 stain but not from the<br />other strain C27-2. The purity of > 99% and recovery of about 40% were obtained as shown by SDS-PAGE<br />and Western blot analysis. The purified r-hGH was biologically active as judged by receptor assay with<br />very low endotoxin content which could be suitable for therapeutic applications. This simple and cost effective<br />production process could be useful for large scale production of recombinant hGH from specific strains.<br /><br />National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001Induction of Spawning in Common carp Cyprinus carpio,Using Pituitary Extract and GnRH Analogue in Combination with Domperidone2132176880ENSalar DorafshanDepartment of Fisheries, College of Natural Resources, Isfahan University of Technology (IUT), Zip code:84154, Isfahan.Hossein MostafaviNational Research Center of Genetic Engineering and Biotechnology (NRCGEB), P.O. Box:14155-6343, Tehran.Bagher Mojazi AmiriDepartment of Fisheries and Environmental Science, Faculty of Natural Resources,
University of Tehran, P.O. Box: 31585-4314, Karaj, I.R. Iran.Journal Article20031001The effectiveness of the first Iranian made gonadotropin releasing hormone analogue, [D-Ala6<br />des-Gly10] GnRH ethylamide, alone or in combination with domperidone, a dopamine antagonist on spawning<br />rate, latency period, working fecundity and embryo viability in common carp, Cyprinus carpio, was investigated.<br />Fifty two fish were divided into 5 groups and treated intrapretoneally as follows: 3 mg/Kg b.w. of carp pituitary extract (C.P.E.) as a positive control, GnRHa alone, 10 μg/Kg b.w.. or in combination with domperidone, 5 mg/Kg b.w. in a single or double injections 7h apart. A group was treated with propylene glycol 0.2 ml/Kg b.w. alone and considered as control. No female ovulated in groups receiving either propylene glycol or 10 μg/Kg b.w. of GnRHa alone. The spawning rate was higher in female GnRHa+domperidone (10 μg/Kg b.w..+5 mg/Kg) in double injections (11 out of 12) as compared to fish which injected either with C.P.E (7 out of 16) or GnRHa + domperidone in a single injection (3 out of 12)(P<0.05). The mean working fecundity, was significantly higher for fish receiving GnRHa+domperidone in single (126214 ±24315) or double injections (145600 ± 27113) compared to C.P.E treated group (52435 ± 1224) (P<0.05). There were no significant differences for latency period or embryo viability among the groups.National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001Initiation of Ageing Process by Meiotic and Mitotic Recombination within the Ribosomal DNA Genes in Saccharomyces cerevisiae2182236885ENMajid Motovali-BashiGenetics group, Biology Department, Faculty of Sciences, Isfahan University, Isfahan, I. R. Iran.Zohreh HojatiGenetics group, Biology Department, Faculty of Sciences, Isfahan University, Isfahan, I. R. Iran.Richard WalmsleyBiomolecular Sciences Department, UMIST, Sackvill St., P.O. Box: 88, Manchester, M60 1QD, UK.Journal Article20031001In the budding yeast of Saccharomyces cerevisiae the tandem repeated of rDNA genes are located on<br />chromosome XII, which is in the nucleolus. There are different types of proteins in the nucleoluskeleton,<br />silencing proteins have got important role in nucleolus.<br />It is shown that meiotic recombination between nonsister chromatids in the rDNA genes are strongly<br />suppressed, and suggested that silencing proteins such as SIR2 are involved in silencing state. It is also<br />shown that nucleolus shows some changes during ageing process. It is claimed that intrachromosomal<br />recombination within the rDNA repeated sequences; producing 3-μm rDNA circles are accumulated with old<br />cell. This study looked at the rDNA breakage in two different types of strain ORD 1181 according to their<br />age. The fine analysis of the rDNA array was performed using restriction endonuclease enzymes, which<br />do not cleave within the rDNA array. The results suggest that there are only meiotic hot regions for chromosome breakage in the old cells within the rDNA array.<br /><br />National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001Sequence-Based Differentiation of Strains in the Streptomyces cyaneus Species-Group2242336887ENEhsan RashidianDepartment of Microbiology, School of Veterinary Medicine, Lorestan University, Khorramabad, I.R. Iran.Michael Goodfellow2School of Biology, University of Newcastle, Newcastle Upon Tyne, NE1 7RU, United Kingdom.Journal Article20031001In an extensive numerical phenetic survey, a number<br />of blue, red and gray spored streptomycete strains<br />were grouped together as Streptomyces cyaneus<br />species-group, a taxon which encompasses strains<br />known to produce antitumor antibiotics, notably anthracyclines.<br />In the present investigation these and related<br />streptomycetes were the subject of morphological and<br />16S rRNA sequencing studies designed to clarify their<br />taxonomic relationships. It is evident from these results<br />that the Streptomyces cyaneus species-group encompasses<br />misclassified strains and members of several<br />distinct species. However, some of the red-spored<br />strains (e.g. Streptomyces janthinus ISP 5206T,<br />Streptomyces roseoviolaceus ISP 5277T and<br />Streptomyces violatus ISP 5209T) formed a distinct<br />clade. In contrast, most of the blue and gray-spored<br />strains showed much less sequence homology<br />between and with one another and were scattered<br />throughout the 16S rRNA Streptomyces tree. The<br />improved classification of this group of streptomycetes<br />provides an essential basis for establishing their industrial<br />and economical significance.<br /><br />National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001A Heterologous Enzyme Linked Immunosorbant Assay of Morphine Using Penicillinase as Label2392466881ENShima HallajFood and Drug Control Laboratories, Department of Biology, Tehran, Iran.Mohammad J. RasaeeDepartment of Biochemistry, Faculty of
Medical Sciences, Tarbiat Modarres University, Tehran, Iran.Monirsadat HaerianFood and Drug Control Laboratories, Department of Biology, Tehran, Iran.Malihe PaknejadDepartment of Biochemistry, Faculty of
Medical Sciences, Tarbiat Modarres University, Tehran, Iran.Soheila KashanianDepartment of Biochemistry, Faculty of
Medical Sciences, Tarbiat Modarres University, Tehran, Iran.Fatemeh RahbarizadehFood and Drug Control Laboratories, Department of Biology, Tehran, Iran.Kobra OmidfarDepartment of Biochemistry, Faculty of
Medical Sciences, Tarbiat Modarres University, Tehran, Iran.Mohammad MalekanehDepartment of Clinical Biochemistry, Birjand University of Medical Sciences, Birjand, Iran.Mohammad Kakhki4Department of Pilot, Pasture Institute of Iran, Tehran, I.R. Iran.Journal Article20031001A rapid, sensitive, specific and high through-put enzyme-linked immunosorbant assay (ELISA) method<br />for determination of morphine in urine samples using penicillinase as label enzyme has been developed. No<br />extraction or chromatography was included in this assay procedure. Immunoglobulin (Ig) purified polyclonal<br />anti-bodies against a C6-hemisuccinate derivative of morphine (M-C6-HS) conjugated to bovine<br />serum albumin (BSA) was coated onto the wells of microtiter plate. A morphine-C3-hemisuccinate (M-C3-<br />HS) was also prepared and the two derivatives were conjugated to penicillinase (M-C6-HS-P and M-C3-HSP).<br />The heterologous combination of antibody prepared against M-C6-HS-BSA and enzyme conjugate<br />prepared for M-C3-HS-P showed better properties in term of sensitivity, reproducibility and slope of standard<br />curve. The assay was sensitive from 20 pg/ml and detected up to 100 ng/ml of morphine in urine samples.<br />The affinity of antibody in homologous assay was found to be 6.6×1010 l/mol and for heterologous assay<br />was 3.2 ×1012 l/mol. The assay was completed within 4 h. The homologous assays performed under different<br />conditions of coating, concentrations, duration, pH, etc. did not end up with a suitable standard curve.<br />Hence it seems that the ability of morphine to displace the hapten enzyme conjugate dependds on the position of the enzyme coupled to the hapten molecule.This ELISA techniqu showed 100% correlation with<br />immunochromatography (IC) and 90% percent correlation with latex agglutination inhibition (LAI) test in the<br />results obtain with urine samples declared positive by authorities. ELISA also showed approximately 90%<br />correlation with LAI-negative urine samples.<br /><br />National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30431420031001Growth and Isolation of Human Cytomegalovirus on a New Human Fetal Foreskin Fibroblast-derived Cell Line in Iran2472516886ENSamad Amini Bavil OlyaeeVirology Department, School of medical Sciences, Tarbiat Modarres University, P.O. Box: 14115-111, Tehran, Iran. Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran.Farzaneh SabahiVirology Department, School of medical Sciences, Tarbiat Modarres University, P.O. Box: 14115-111, Tehran, Iran.Mohammad Hassan RostaieVirology Department, School of medical Sciences, Tarbiat Modarres University, P.O. Box: 14115-111, Tehran, Iran.Mohsen Karimi ArzenaniBiotechnology Department, Pasteur Institute of Iran, Tehran, Iran.Zahra Samadi Bahrami3National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, I.R. Iran.Ramin Sarrami ForooshaniBiotechnology Department, Pasteur Institute of Iran, Tehran,l Iran.Ahmad AdeliBiotechnology Department, Pasteur Institute of Iran, Tehran, Iran.Journal Article20031001Cell culture technique has been used for detection and confirmation of many different viruses in clinical samples. Although, new diagnostic methods have been developed for viral infections, traditional cell culture<br />technique is still regarded as the “gold standard” for several infectious agents such as human cytomegalovirus<br />(HCMV). In the present study, a new human fetal foreskin fibroblast (HFFF)-derived cell line was obtained from a normal Iranian male fetus at the end of first trimester at National Cell Bank of Iran (NCBI) and its ability for HCMV growth was investigated.<br />Thirteen treated urine samples from renal transplant recipients were inoculated onto HFFF monolayer.<br />HCMV growth was monitored and confirmed by typical CMV cytopathic effect (CPE), histological staining,<br />immunological staining and polymerase chain reaction (PCR)-based restriction fragment length polymorphism<br />(RFLP) method and sequencing. All of the abovementioned<br />detection methods were confirmed that new<br />HFFF-derived cell line is a sensitive line for growth of<br />HCMV. This cell line has been named HFFF-PI6 and is<br />available with accession number NCBI C170 at NCBI<br />of Pasteur Institute of Iran.<br /><br />