摘要
从人 18周胎脑中抽提mRNA ,建立cDNA文库 ,通过大规模测序克隆出一条人类基因 ,该基因全长16 44bp ,开放阅读框 1146bp ,编码一个含 372个氨基酸的蛋白 ,预测分子质量约为 46 8ku .经与蛋白数据库比较 ,发现具有人硫氧还蛋白结构 ,命名为人的类硫氧还蛋白基因Ⅰ (humanThioredoxinlikegeneⅠ ,hTRXLⅠ ) .多组织膜Northernblot结果显示在心脏、脑、胎盘、肝、骨骼肌、肾、胰腺等各个组织中均有表达 ,肺中表达不明显 .通过Unigene染色体定位 ,将hTRXLⅠ定位于 11q11,把hTRXLⅠ的读框克隆入经改造的PBV2 2 0质粒中 ,在E .coli中诱导后获得表达 .
Through screening a human fetal brain (18 weeks) cDNA library, a cDNA was isolated. It's 1 644 bp in full length and ORF was 1 146 bp. The putative protein had 372 amino acids, with molecular weight about 46.8 ku. Compared with that of protein data pool, the putative protein was found to have human thioredoxin structure. So it was named human thioredoxin like gene Ⅰ ( hTRXL Ⅰ). Northern blot analysis of multiple tissues indicated hTRXL Ⅰ expressed in heart, brain, placenta, liver, skeletal muscle, kidney, pancreas and so on, but not in lung. By using Unigene, hTRXL was mapping to human chromosome 11q11. The ORF of hTRXL Ⅰ was subcloned into reconstructed PBV220 plasmid, and found expression in BL21 with IPTG inducing.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
2000年第6期593-597,共5页
Journal of Fudan University：Natural Science
基金
上海市现代生物与新药产业发展基金资助项目!(98431912 1)
