Molecular and Bioinformatics Analysis of Allelic Diversity in IGFBP2 Gene Promoter in Indigenous Makuee and Lori-Bakhtiari Sheep Breeds
محورهای موضوعی : Camelع. ولی‌پور کوتنایی 1 , ا. فرهادی 2 , س.ح. حافظیان 3 , م. قلی‌زاده 4
1 - Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
2 - Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
3 - Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
4 - Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
کلید واژه: promoter, sequencing, Lori-Bakhtiari sheep, Makuee sheep, <i>IGFBP gene</i>,
چکیده مقاله :
The aim of this study was to perform molecular and bioinformatics analysis of IGFBP2 gene promoter in association with some economic traits in indigenous Makuee (MS) and Lori-Bakhtiari (LB) breeds. DNA was extracted from blood samples of 120 MS and 200 LB and a 297 bp fragment from the upstream sequences of studied gene was amplified and genotyped by single-strand conformational polymorphism (SSCP) technique. Two genotypes of AB and BB were seen in MS and LB breeds. Then one sample from each genotype was send to sequencing. After obtaining the sequencing result, the sequences homology was performed on the National Center for Biological Information NCBI server by basic local alignment search tool (BLAST) program. The alignment of the obtained sequences and their comparison with reference sequences from the Gene Bank were done using CLUSTALW multiple alignment tool of BioEdit software. In addition, the DNASIS MAX software was used to identify DNA motifs. The bioinformatics analysis revealed differences in sequences of IGFBP2 between observed genotypes. Ten motifs in promoter sequence of IGFBP2 genes were seen, so that the CAP_site motif was most abundant in both fragments motif. Statistical analysis using general linear method model (GLM) procedure of SAS software showed significant (P<0.05) association of IGFBP2 with thigh round (TR) trait in Makuee sheep. Further studies in other indigenous sheep breeds and investigation of other genetic regions along with regulatory sites seem to be necessary.
هدف از پژوهش حاضر تجزیه و تحلیل مولکولی و بیوانفورماتیک تنوع آللی پروموتر ژن IGFBP2 در ارتباط با برخی صفات مهم اقتصادی در گوسفندان بومی نژادهای ماکویی و لری بختیاری بوده است. نمونههای DNA از خون 120 رأس گوسفند ماکویی و 200 رأس گوسفند لری بختیاری استخراج و یک توالی 297 جفت بازی مربوط به ناحیه بالادست ژن مورد مطالعه تکثیر و با تکنیک SSCP تعیین ژنوتیپ شد. دو ژنوتیپ AB و BB در نژادهای ماکویی و لری بختیاری مشاهده شدند. سپس از هر ژنوتیپ یک نمونه برای تعیین توالی ارسال شد. پس از دریافت نتایج توالییابی، بررسی همولوژی توالیها با برنامه BLAST روی سایت NCBI انجام شد. همترازی توالیهای بدست آمده و مقایسه آنها با توالی رفرنس از بانک ژنیبا ابزار با CLUSTALW نرمافزار Bio Edit صورت گرفت. علاوه بر این، شناسایی موتیفهای DNA با استفاده از نرمافزار DNASIS MAX انجام شد. بررسیهای بیوانفورماتیک تفاوتهایی را بین توالی ژنوتیپهای مختلف IGFBP2 نشان داد. 10 موتیف در توالی پروموتر ژن IGFBP2 مشاهده شد، به طوری که بیشترین فراوانی مربوط به موتیف CAP_site بود. بررسی ارتباط آماری با پروسه GLM نرمافزار آماری SAS وجود ارتباط معنیدار آماری (05/0P<) بین پروموتر ژن IGFBP2 و صفت دور ران (TR) در گوسفند ماکویی را نشان داد. انجام پژوهش‌های بیشتر در سایر نژادهای بومی و بررسی سایر نواحی ژنی همراه با نواحی تنظیمی این جایگاه پیشنهاد میشود.
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