Title: Cloning and Bioinformatics Analysis IGF-I Gene in Pig
Abstract: In order to clone the pig insulin-like growth factor I( IGF-I) and detect the transcription level of the factor in different tissues,the RNA of 13 kinds of tissues was extracted,then the RNA was subjected to reverse transcription to form cDNA,and the gene encoding region entire sequence of the IGF-I was amplified through a RT-PCR method,then the purified product of PCR was ligated with a pMD18-T and transferred into the bacterium DH5α for replication. The positive clones were screened and sequenced. The sequence character analyzed by using bioinformatics method and phylogeny evolution tree was constructed with other five species,such as cattle. Meanwhile,the semi-quantitative RT-PCR was used to analyze the expression profile of IGF-I gene in 13 kinds tissues. The result showed the coding region of IGF-I gene was 462 bp( GenBank accession number: JX827417),and coded 153 amino acids. The prediction of secondary structure indicated that random coils were percent 57. 52. The amino acid homology analysis showed that pig IGF-I had high homology with the IGF-I of five species:cattle( 98. 0 %),horse( 97. 4 %),sheep( 96. 1 %),deer( 96. 1 %),chicken( 83 %). Semi-quantitative RT-PCR analysis of mRNA from different tissues indicted that IGF-I mRNA expressed in 13 tissues,and the expression level was higher in muscle,liver,spleen,ovary and large intestine than other tissues.
Publication Year: 2014
Publication Date: 2014-01-01
Language: en
Type: article
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