Cloning, Expression and Bioinformatics Analysis of IGF-1 and IGF-2 Genes in Congjiangxiang Pigs (Sus scrofa)
ZHU Xiao-Feng1,2, XU Hou-Qiang1,2,*, CHEN Wei1,2,3, CHEN Ying-Lian1,3, NI Kai1,2, WU Xiao-Min1,2, NI Meng-Meng1,2, LU Xian-Jun1,2
1 Key Laboratory of Animal Genetics, Breeding and Reproduction in the Platean Mountainous Region Ministry of Education/Key Laboratory of Animal Genetics, Breeding and Reproduction, Guizhou University, Guiyang 550025, China; 2 College of Animal Science, Guizhou University, Guiyang 550025, China; 3 College of Life Science, Guizhou University, Guiyang 550025, China
Abstract：Congjiang Xiang pig (Sus scrofa) has the characters of short and small body and slow growth rate. For preliminarial exploreation on Congjiang Xiang pig's characters and providing basic data on the molecular mechanism of growth and development of miniature pigs, insulin-like growth factor 1 and 2 (IGF-1 and IGF-2) genes had been analysed by bioinformatics methods as well as their expression level of mRNA in various organs and tiusses from Congjiang Xiang pig and Norther Yorkshinre had been detected. In the current, the coding sequences (CDS) of IGF-1 (GenBank No. NM_214256.1) and IGF-2 (GenBank No. NM_213883.2) genes were had successfully cloned by using Congjiang Xiang pig and Norther Yorkshinre with same feeding condictions and ages and had predicted and dissected on the structures and functions of IGF-1 and IGF-2. The qRT-PCR had been applied to detect mRNA levels of IGF-1 and IGF-2 of Congjiang Xiang pig and North yorkshire in the heart, liver, spleen, lung, kidney, large intestine, small intestine, fat, and longissimus 9 organs or tissues. The results showed that the CDS of IGF-1 and IGF-2 of Congjiang Xiang pig were 435 and 577 bp respectively, which were no difference with CDS sequence of Sus scrofa in GenBank. By sequence prediction, the relative molecular mass, theoretical isoelectric point, and unstable coefficient of IGF-1 and IGF-2 protein resplectively were 14.390 44 and 20.312 5 kD, 9.34 and 10.00, 65.58 and 58.64. Comparing other 12 species by genetic evolution analysis showed that Congjiang Xiang pig was the closest with S. scrofa. The qRT-PCR results displayed that IGF-1 gene had the obviously highest expression level in lung, large intestine, and small intestine of Congjiang Xiang pig (P<0.05) as well as IGF-1 gene had the very significantly highest expression level in longissimus of North Yorkshine (P<0.01) by comparing other organs or tissues. The IGF-2 had obviously higher expression level in liver and lung than other organs and tissues in Congjiang Xiang pig but longissimus and heart of North yorkshine had very significantly higher expression level of IGF-2 gene than other organs or tissues (P<0.01). Our research uncovered the expression difference of IGF-1 and IGF-2 genes between Congjiang Xiang pig and North Yorkshine, which provided basic data for further exploration of the molecular mechanism of growth and development of miniature pigs.
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