بررسی چندشکلی در اینترون سوم ژن هورمون رشد گاو بومی گیلان با استفاده از روش PCR-RFLP

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش آموخته کارشناسی ارشد گروه علوم دامی دانشکده علوم کشاورزی دانشگاه گیلان

2 دانشیار گروه علوم دامی دانشکده علوم کشاورزی دانشگاه گیلان

3 دانشیار گروه زراعت و اصلاح نباتات دانشکده علوم کشاورزی دانشگاه گیلان

4 عضو هیات علمی موسسه تحقیقات برنج کشور- رشت

چکیده

ژن هورمون رشد گاوی (bGH)، بخشی از خانواده ژن­های پرولاکتین و لاکتوژن­های جفتی است. تنوع موجود در اینترون‌های این ژن، پتانسیل ویژه­ای برای استفاده به­عنوان مارکرهای ژنتیکی از خود نشان داده و می­تواند به بهبود ژنتیکی جمعیت­ها کمک کند. به­منظور بررسی فراوانی آللیMspI در اینترون سوم ژن هورمون رشد گاوهای بومی گیلان، به­طور تصادفی 70 رأس گاو از این جمعیت تعیین و از آنها نمونۀ خون تهیه شد. با استفاده از روش تغییر یافته استخراج نمکی، DNA نمونه­ها استخراج شد. سپس با استفاده از روش واکنش زنجیره­ای پلیمراز (PCR) قطعۀ 345 جفت بازی در این ناحیه تکثیر و محصول PCR با استفاده از آنزیم MspI هضم شد. سپس به­منظور تعیین ژنوتیپ، نمونه­های هضم شده به ژل اکریل آمید 8% منتقل شدند. این آنزیم دارای 2 مکان برش روی قطعه مورد نظر بوده و در اثر هضم با آنزیم MspI سه ژنوتیپ مشاهده شده و فراوانی آللی (+)MspI 4643/0 محاسبه شد. آزمون کای مربع بیانگر وجود حالت تعادل در فراوانی محاسبه شده برای آلل MspI بود.

کلیدواژه‌ها


عنوان مقاله [English]

Polymorphisms in third intron of growth hormone of Guilan native cows using PCR-RFLP markers

نویسندگان [English]

  • R. Seighalani 1
  • S. Z. Mirhosseini 2
  • B. Rabiei 3
  • A. A. Ebadi 4
1 Graduated MS student, Animal Science Department, University of Guilan
2 Associate Professor, Animal Science Department, University of Guilan
3 Associate Professor, Plant Breeding Department, University of Guilan
4 Researcher, Rice Research Institute of Iran
چکیده [English]

Bovine growth hormone(bGH) gene, is a part of the multiple gene family that contains prolactin and
placental lactogens. The variations in the introns of this gene, have potential usefulness as genetic markers and
could help in the genetic improvement of populations. In ordre to investigate allele frequency of MspI in introne
3 of growth hormone gene of Guilan native cattle breed, blood samples were collected from seventy heads of
randomaly choosen cattles (cows). Genomic DNA was extracted from these samples, using modified salting out
method. Polymerase chain-reaction (PCR) procedure was used to amplify a 345 bp segment in this region and
then digested with MspI restriction enzymes. Digested samples were run on the acrylamid gel 8% to recognize
their genotype. This enzyme had two restriction sites and after digestion with MspI enzyme on that fragment and
2 allels and 3 genotypes have been observed. Frequency of MspI(+) allele was estimated to 0.4643. X2 test has
showed the equilibrium of frequency MspI in samples.

کلیدواژه‌ها [English]

  • Bovine growth hormone
  • DNA polymorphism
  • Guilan native cow
  • MspI allelic frequency
  • Third intron
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