National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Pneumoviruses: Molecular Genetics and Reverse Genetics78936944ENGholamreza AhmadianDepartment of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, P.O. Box:
14155-6343, Tehran, I.R. IranMehdi ShamsaraDepartment of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, P.O. Box: 14155-6343, Tehran, I.R. Iran. Department of Genetics, Faculty of Basic Sciences, Tarbiat Modarres University, P.O. Box: 14115-111,Tehran, I.R. Iran.Andrew J. EastonDepartment of Genetics, Faculty of Basic Sciences, Tarbiat Modarres University, P.O. Box: 14115-111,Tehran, I.R. Iran.Journal Article20050401Pneumoviruses are responsible for significant respiratory disease in their hosts and represent a major problem<br />for human and animal health. Pneumoviruses are members of the family Paramyxoviridae, subfamily<br />Pneumovirinae and the virus particles consist of a negative-sense, nonsegmented RNA genome within a helical nucleocapsid structure enveloped in a lipid membrane derived from the host cell. Over the past four<br />decades much work has extended our understanding of the molecular biology and pathogenesis of pneumoviruses but despite this only limited treatments and prophylaxis are available. The human pathogen, respiratory syncytial virus (hRSV) which belongs to the genus of Pneumovirus is the best characterized of the<br />subfamily. HRSV is the major cause of hospitalisation of very young children with respiratory disease worldwide. No vaccine is available though new treatments offer some respite for children in the highest risk<br />groups, the immunocompromised and children with congenital heart disease. The recently discovered<br />human pathogen human metapneumovirus (hMPV) belongs to the genus Metapneumovirus and recent<br />data indicates that this virus is second only to hRSV in terms of disease impact. The pneumoviruses also<br />include agents of veterinary importance such as bovine respiratory syncytial virus (bRSV), ovine and<br />caprine RSV, and pneumonia virus of mice (PVM: all in the genus Pneumovirus) and avian metapneumovirus<br />(APV: genus Metapneumovirus). The development of reverse genetics systems for negative strand RNA<br />viruses has opened the possibility of manipulating the virus genomes to identify genes involved in pathogenesis and to explore the biological consequences of specific mutations. This information is informing the rational design of new vaccines. These plasmid-based systems have shown that for all paramyxoviruses the N, P and L proteins are necessary and sufficient for RNA replication. However, the pneumoviruses differ<br />from the other family members in that fully efficient transcription from the virus genome requires the presence<br />of an additional protein encoded by the M2 gene. The present article reviews pneumovirus biology and<br />molecular genetics including a discussion of current concepts of Pneumovirus reverse genetics.<br /><br />https://www.ijbiotech.com/article_6944_786686d44e2c0135b7c4ed989a7c4e9c.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Specific Inhibition of the Expression of the Promyelocytic Leukemia (PML) Protein by Anti-Sense Oligonucleotides94986959ENSadeq VallianDivision of Genetics, Department of Biology, Faculty of Science, Isfahan University, Isfahan, I.R. IranKun-Sang ChangDepartment of Laboratory Medicine, MD Anderson Cancer Center, Houston, Texas, USA.Journal Article20050401In the present study, using anti-sense oligonucleotides the inhibition of expression of the PML protein has<br />been investigated. The anti-sense oligonucleotides were designed against the translation initiation site of<br />the PML gene, and their effects were investigated on cellular growth and DNA synthesis. Incubation of normal<br />human fibroblast cells with the anti-sense oligonucleotides resulted in the complete inhibition of the PML<br />protein expression. Inhibition of the PML protein expression by anti-sense oligonucleotides was found<br />to be associated with an increase in cellular growth and doubling time. Furthermore, in cells treated with<br />the anti-sense oligonucleotides, but not sense or scrambled oligonucleotides (control), the cellular DNA<br />synthesis also showed a marked increase, confirming the induction of cellular growth upon inhibition of PML<br />synthesis. These findings clearly demonstrated that the inhibition of the expression of the PML protein could be achieved using the anti-sense oligonucleotides, providing a model for better investigation of the biologic role of PML in the cell.<br /><br />https://www.ijbiotech.com/article_6959_f4e1b5843a1d251d0b434225523b3776.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Molecular Typing of Avian Pasteurella multocida lsolatesby PCR-RFLPof ompH Gene991036964ENAhmad Reza JabbariDepartment of Aerobic Veterinary Bacterial Vaccines, Razi Vaccine and Serum Research Institute, Hesarak, Karaj, I.R. IranMajid EsmaelizadehDepartment of Biotechnology, Razi Vaccine and Serum Research Institute, Hesarak, Karaj, I.R. Iran.Journal Article20050401Molecular typing of twenty-five Pasteurella multocida isolates has been assessed by restriction fragment<br />length polymorphism (RFLP) of a species-specific PCR assay. Amplification was based on the gene ompH, encoding a major outer memberane protein. RFLP analysis of the 1.2 kb ompH-amplification using<br />EcoRI, HindIII and CfoI endonucleases produced 7 different patterns for the twenty five isolates of the four<br />P.multocida serotypes. The PCR-RFLP of the ompH gene was found to be potentially a useful method for<br />typing of P. multocida and therefore, for studying the epidemiology of P. multocida infections.<br /><br />https://www.ijbiotech.com/article_6964_ac6dc1beb7cd8200f3010e50680c76b1.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401The Allele and Genotype Frequencies of Bovine Pituitaryspecific Transcription Factor and Leptin Genes in IranianCattle and Buffalo Populations Using PCR-RFLP1041086943ENArash JavanmardDepartment of Genomics, West and North-West Agriculture Biotechnology Research Institute(ABRII-T), Tabriz, I.R. Iran.Nader AsadzadehDepartment of Animal Production and Management, Animal Science Research Institute of Iran (ASRI), Karaj, I.R. Iran.Mohammad Hossein BanabaziDepartment of Biotechnology, Animal Science Research Institute of Iran (ASRI),
Karaj, I.R. Iran.Javad TavakolianDepartment of Biotechnology, Animal Science Research Institute of Iran (ASRI),
Karaj, I.R. Iran.Journal Article20050401The use of polymorphic markers in breeding programmes could make selection more accurate and efficient. A total of 324 individuals from six Iranian cattle populations (Sarabi, Golpayegani, Sistani, Taleshi, Mazandarani, Dashtiyari), F1 Golpayegani × Brown Swiss and Iranian buffalo populations were genotyped<br />for the Pit-1 HinfI and leptin Sau3AI polymorphisms by the polymerase chain reaction and restriction fragment<br />length polymorphism (PCR-RFLP). The genotype and gene frequencies for each breed were determined and<br />shown to be quite variable among the breeds. The highest frequencies of allele B for the leptin gene and<br />allele A for the Pit-1 gene were found in Dashtiyari and Sistani cattle, respectively. According to our results,<br />the highest AB genotype frequencies were found in the Taleshi and F1 Golpayegani x Brown Swiss cross for<br />the leptin and Pit-1 genes, respectively. These allele frequencies were comparable to previously published<br />data on exotic breeds. The highest and lowest heterozygosities were found in Taleshi and Dashtiyari cattle<br />for the leptin gene and in F1 Golpayegani x Brown Swiss cross and Sistani cattle for the Pit-1 gene, respectively. These values indicated the presence of low variation for these genes in the studied populations.<br />The possible association between molecular polymorphisms within these candidate genes and economic<br />traits for the studied populations should be further investigated.<br /><br />https://www.ijbiotech.com/article_6943_10d06165d9d69e766305ae15cbcd593a.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Transient expression of human growth hormone in potato (Solanum tuberosum), tobacco (Nicotiana tobacum) and lettuce (Lactuca sativa) leaves by agroinfiltration1091136965ENHaleh Hashemi SohiDepartment of Plant Molecular Biology, National Institute for Genetic Engineering and Biotechnology (NIGEB), P.O. Box: 14155-6343, Tehran, I.R. IranEsmat JourabchiDepartment of Plant Molecular Biology, National Institute for Genetic Engineering and Biotechnology (NIGEB), P.O. Box: 14155-6343, Tehran, I.R. Iran.Mahvash KhodabandehDepartment of Bioprocess Engineering, National Institute for Genetic Engineering and Biotechnology (NIGEB), P.O. Box: 14155-6343, Tehran, I.R. IranJournal Article20050401Using agro-infiltration technique, we have transiently expressed human Growth Hormone (hGH) in tobacco<br />(Nicotiana tobacum), potato (Solanum tuberosum) and lettuce (Lactuca sativa) leaves. Out of three different<br />inoculation times used for infiltration in our study, it was seen that highest level of hGH expression was<br />achieved when leaves were infiltrated for 35 min. The presence of biologically active hGH was detected in<br />leaf extract by western blotting and ELISA. The highest expression was measured in tobacco was found to<br />be1.5-3 hGH mg/kg leaves.<br /><br />https://www.ijbiotech.com/article_6965_19c083330a245a15813be5fc5840dcbd.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Growth improvements of Sunflower seedlings by Cr(VI)-resistant bacteria1141206963ENMuhammad FaisalDepartment of Botany, Faculty of Agriculture, University of the Punjab, Quaid-e-Azam Campus, Lahore-54590, Pakistan.Shahida HasnainDepartment of Botany, Faculty of Agriculture, University of the Punjab, Quaid-e-Azam Campus, Lahore-54590, Pakistan.Journal Article20050401In the present study, three chromium resistant bacterial strains (CrT-1, CrT-2, CrT-3) which could resist very<br />high concentration of K2CrO4 (up to 40 mg ml-1 on nutrient agar plates and 10 mg ml-1 in acetate-minimal<br />medium) were used to inoculate the sunflower seeds both as control and under chromium stress. Cr(VI)<br />caused severe reduction in different growth parameters (seedling length, fresh weight, dry weight g-1 fresh<br />weight) as compared to control, while bacterial inoculations improved different growth parameters both as<br />control and under chromate stress when compared with non-inoculated respective controls. With respect<br />to biochemical parameters, acid phosphatase and auxin content showed marked increment with bacterial<br />inoculation both in chromium stress and unstressed condition. Uptake of chromium in inoculated plants<br />decreased significantly as compared to non-inoculated control. Cr (VI) application also severely damages different plant cells/tissues but bacterial inoculation not only improves the growth and yields parameters but<br />also prevent cell damages caused by the Cr (VI) salt.<br /><br />https://www.ijbiotech.com/article_6963_4c32bbaeb338e4568da96f9095503812.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Transfection of 6-23 a rat thyroid carcinoma cell line, with the Neuropeptide Y cDNA1211246945ENMehdi MahmoodiDepartment of Biochemistry and Biophysic, Faculty of Medicine, Rafsanjan, I.R. Iran.0000-0002-8463-8364Hamid Reza RashidinejadDepartment Internal Medicine, faculty of Medicine, Rafsanjan, I.R. Iran.Gholamreza Asadi KaramDepartment of Biochemistry and Biophysic, Faculty of Medicine, Rafsanjan, I.R. Iran.Mohammad KhaksariDepartment of Physiology and Pharmacology, Faculty of Medicine, Kerman, I.R. Iran.Ebrahim MirzajaniDepartment of Biochemistry, Faculty of Medicine, Rasht, I.R. Iran.Stephen Robert BloomDepartment of Metabolic Medicine, Imperial College School of Medicine, London, England.Journal Article20050401Neuropeptide Y (NPY) is a 36 amino acid peptide found throughout the central and peripheral nervous<br />system of rat and human. NPY has been proposed to play an important role in satiety. The aim of this study<br />was to produce cell lines that secrete high levels of bioactive NPY. For this purpose, the complementary<br />DNA (cDNA) that encodes NPY was isolated by PCR. The cDNA was then cloned into pCEP4, to form<br />pCEP4NPY. 6-23 cells were transfected with pCEP4NPY by electroporation. Transfected cells were selected by the addition of hygromycin B to the culture medium. Resistant colonies were picked and transferred to 96-well plates. The medium was tested for IRNPY using a specific NPY radioimmunoassay (RIA). The IR-NPY secreted by the cells was characterized by sephadex G50 chromatography and reversed phase fast protein liquid chromatography (FPLC). It was found to co-elute with the synthetic standard in both cases. RNA was extracted from the cells and subjected to Northern blot analysis using labeled NPY cDNA as a probe. The cells were found to express high levels of NPY at mRNA levels.https://www.ijbiotech.com/article_6945_dc66ef03fd8577f8377794647a116b3e.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30433220050401Investigation of two recessive disorders in breeder bulls of Abbas Abad animal breeding center1251286948ENMohammad Reza NassiryAnimal Science Department, College of Agriculture, Ferdowsi University of Mashhad, P.O. Box: 91775-1163
Mashhad, I.R. Iran.Amir NorouzyAnimal Science Department, College of Agriculture, Ferdowsi University of Mashhad, P.O. Box: 91775-1163
Mashhad, I.R. Iran.Fereidoun Eftekhari ShahroudiAnimal Science Department, College of Agriculture, Ferdowsi University of Mashhad, P.O. Box: 91775-1163
Mashhad, I.R. Iran.Ali JavadmaneshAnimal Science Department, College of Agriculture, Ferdowsi University of Mashhad, P.O. Box: 91775-1163
Mashhad, I.R. Iran.Mohammad Ali ShadCenter of Animal Breeding in North-Eastern of Iran, Jahad-Keshavarzi Khorasan, Mashhad, I.R. IranJournal Article20050401To prevent distribution of recessive alleles in dairy herds all bulls used for AI (Artificial Insemination) have<br />to be tested. In this study 26 blood and 4 semen samples were supplied from Iranian Holstein bulls used for<br />AI. Genomic DNA was extracted from 100 μl of blood and 200 μl of semen. Samples were tested by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method. PCR reaction was performed for amplification of polymorphic region of the CD18 gene on chromosome 1 and exon 5 of ASS gene on chromosome 11. We detected one Bovine Leukocyte Adhesion Deficiency (BLAD) carrier and no carrier for bovine Citrolliemia in this study. Hardy-Weinberg test confirmed the equilibrium of BLAD locus in this population.https://www.ijbiotech.com/article_6948_4a4dd5c487a083e34bea1d68807d018e.pdf