牛PS ME1基因的电子克隆与序列分析

In silico cloning and characterization of bovine PS ME1 gene

电子克隆是基因克隆的新策略.以人类PSME1基因完整编码序列（NM_006263）为种子序列电子克隆了牛PSME1基因完整编码序列.以奶牛外周血白细胞cDNA为模板,根据电子克隆序列设计引物进行RT-PCR验证,PCR产物克隆入质粒载体,阳性克隆测序得到的序列与电子克隆序列一致,已被GenBank收录,序列号为DQ010410.该基因最大开放阅读框34~783 bp,编码249个氨基酸,预测其编码的蛋白分子量为28.66 kD,等电点5.87.

In silico cloning was a new strategy of gene cloning which developed with the development of genome, EST projects and bioinformatics. The complete coding sequence of bovine PSME1 gene was obtained by using in silico cloning. The in silico cloned cDNA sequence has been verified by RT-PCR from the single-strand cDNA which was synthesized from total RNA of peripheral blood leukocyte. The cDNA was 883 bp,contains a 750 bp（34~783 bp） open reading frame,encoding 249aa protein,this putative protein weight molecular was 28.66 kD,isoelectric point was 5.87. The sequence has been submitted to GenBank with the accession number DQ010410.