CDNA Library Construction For Antler Epithelial Tissue Of Cervus Elaphus And Insulin-Like Growth Factor Genes Cloning

Total RNA was extracted from antler epithelial tissue of Cervus elaphus using SV Total RNA Isolation System.The antler epithelial tissue cDNA library of Cervus elaphus was constructed using SMART technique.The library contains 1.81×10~6 pfu/ml clones and the amplified library has a titer of 2.4×10~8 pfu/ml,in which 85%clones are recombinant and the average sizes of inserted cDNAs are about 1.0 kb.These results show that the antler cDNA library is qualified for screening genes.Primers designed according to published literatures and the gene sequences of other mammals were used to amplify the full sequence of IGF1 gene for the first time.The results showed,the full length of IGF1 gene with 469 bp.The comparison of these sequences with those of other animals through Blast of GenBank,the results indicated that the identities sequence of the IGF1 gene were 97.8%,97.6%,93.1%,91.8%and 87.2%by homologous comparison among Cervus elaphus and other five species(Bos taurus,Ovis aries, Sus scrofa,Homo sapiens and Mus musculus),and that were 97.4%,96.8%,94.8%,94.2%and 93.0%in amino acid sequences.The results showed that the IGF1 gene was conservative in the process of evolution.Primers designed according to published literatures and the gene sequences of other mammals were used to amplify the full sequence of IGF2 gene for the first time.The results showed,the full length of IGF2 gene with 540 bp.The comparison of these sequences with those of other animals through Blast of GenBank,the results indicated that the identities sequence of the IGF1 gene were 97.8%,97.6%,93.1%,91.8%and 87.2%by homologous comparison among Cervus elaphus and other five species(Bos taurus,Ovis aries, Sus scrofa,Homo sapiens and Mus musculus),and that were 86.5%,95.8%,95.0%,85.8%,82.3%and 82.7%in amino acid sequences.The results showed that the IGF2 gene was conservative in the process of evolution,but in the DNA level or in some variationThe success of cDNA library from antler epithelial tissue of Cervus elaphus with convertingλTriplEx2 SMART technique and antler epithelial tissue IGFs gene cloning,not only Cervus elaphus growth and development of a clear function of the specific gene cloning, but also fine Antler breeding and gene expression and function of the cloning research laid the material foundation.