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Iranian Journal of Biotechnology، جلد ۷، شماره ۴، صفحات ۲۵۸-۲۶۱
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عنوان انگلیسی Isolation of the Gene Coding for Movement Protein from Grapevine Fanleaf Virus
چکیده انگلیسی مقاله A pair of degenerate primers, GMPF1 and GMPR1, was designed on the basis of alignment of previously reported Grapevine fanleaf virus (GFLV) movement protein (MP) nucleotide sequences from Iran and other parts of the world. cDNA was synthesized by the use of Oligo d(T)18 from total RNA extraction from each diseased grapevine leaf sample and subjected to polymerase chain reaction (PCR) with the degenerate primers under a range of annealing temperatures from 48 to 62°C. It was revealed that 55°C gave the best result in terms of producing exactly the expected fragment (1044 bp) from as many samples as possible although accompanied by few fade non specific fragments. However, by application of “hot-start” PCR and annealing at 60°C the specific fragment was amplified from 41 out of 86 samples. This was the first amplification of the precise MP cDNA from GFLVs in Iran which is very important as to preparation of recombinant anti-GFLV MP antibody to use in studying the GFLV- grapevine interaction, and also for generating pathogen-derived resistant vines.
کلیدواژه‌های انگلیسی مقاله Annealing,degenerate,GFLV,MP,Hot-start,PCR
نویسندگان مقاله نعمت سخندان بشیر | sokhandan bashir
department of biochemistry and molecular biology, oklahoma state university, stillwater, ok 74078, usa

افسانه دلپسند خبازی | delpasand khabbazi
department of plant protection, faculty of agriculture, university of tabriz, p.o. box 51664, tabriz, i.r. iran
سازمان اصلی تایید شده: دانشگاه تبریز (Tabriz university)

اسماعیل ترابی |
department of plant protection, faculty of agriculture, university of tabriz, p.o. box 51664, tabriz, i.r. ira
سازمان اصلی تایید شده: دانشگاه تبریز (Tabriz university)

نشانی اینترنتی http://www.ijbiotech.com/article_7088_93de595966edf639c544af32ce3fad1f.pdf
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