猪色素上皮衍生因子的电子克隆及序列分析

用鼠的色素上皮衍生因子cDNA序列在猪的ESTs库进行BLASTn搜索,得到一系列不同大小的ESTs片段,经拼接得到完整cDNA序列.序列分析显示该cDNA长1 425 bp,有一个1 242 bp的开放阅读框,编码413个氨基酸,5’非编码区长53 bp,3’非编码区长130 bp,有一个加尾序列和多聚腺苷酸尾巴.其核苷酸序列与牛、人和鼠的同源性分别为89%、87%和82%,氨基酸的同源性分别为89%、87%和84%,且有保守的糖基化位点、半胱氨酸位点和serp in基序,说明所克隆的cDNA序列为猪的PEDF全长cDNA.

In Solico Cloning and Sequence Analysis of Pigment Epithelium-derived Factor from Pig, Sus scrofa

Many pig EST sequences were aligned using mouse PEDF cDNA sequence as a probe from pig EST database in NCBI web set,and a full-length cDNA sequence was obtained by splicing the overlapped EST.Sequence analysis showed that the in silico cloned cDNA is 1 425 base pairs long with an open reading frame(ORF) containing 1 242 nucleotides encoding a protein of 413 amino acids.5'-untranslated region(UTR) is 53 bp,and 3'-UTR is 130 bp containing a poly(A) signals(AATAAA) and a poly(A) tail.A comparison of the pig cDNA ORF with cow,human and mouse PEDF ORF nucleotide has 89 %,87% and 82% identity,respectively,and it has 89 %,87 % and 82% identity as to amino acid.The N-glycosylated sites,cysteine residue and serpin motif is conserved by multiple alignment.All these results indicated that the in silico cloned cDNA is pig PEDF gene and this cDNA is a base for molecular cloning of pig PEDF gene and its further research.