人类一条新的类蛋白质二硫键异构酶cDNA的克隆和表达

Cloning Expression of a Novel Human Protein-Disulfide Isomerase Like cDNA

利用 SMART技术构建了人胎脑 c DNA文库 ,通过大规模测序筛选出一条 16 45 bp的人类 c DNA。此c DNA含有一个 888bp的开放阅读框 (ORF) ,编码一个 2 96个氨基酸的蛋白质 ,预测分子量 34 .0 KD。与目前数据库中序列比较 ,该 c DNA编码的蛋白质与蛋白质二硫键异构酶的同源性达 36 % ,命名为类蛋白质二硫键异构酶(PDI- L)基因。多组织 Northern blot分析显示 PDI- L c DNA在心、脑、肝、肾等组织中均有表达。该 c DNA的读框片段正确插入到改造后的 p BV2 2 0表达载体中 。

Using the large scale sequencing, a novel human cDNA of 1 645 bp was screened from the cDNA library of human fetal brain. The cDNA contains an ORF encoding a 296-aa protein with a calculated molecular weight of 34.0 KD. Compared with that of current sequence databases, the putative protein was found to have 36% homology with protein-disulfide isomerase (PDI). So this cDNA was named PDI like (PDI-L) gene. Multiple tissue Northern blot analysis shows that PDI-L cDNA is expressed in heart, brain, liver, kidneys and so on. The ORF fragment of PDI-L cDNA was inserted into reconstructed pBV220 expression vehicle and its predicted expressive protein was obtained.