摘要
人在早期胚胎时ｔＰＡ基因表达，但ｍＲＮＡ丰度很低。以１７日龄人胚胎为材料，分离ｍＲＮＡ，以表达质粒ｐＳＰＯＲＴ１为载体，建立ｃＤＮＡ文库，重组子达到１．６×１０６个，能包含极低表达基因的ＣＤＮＡ。从人血白细胞基因组中扩增出一条１７５ｂｐ的ｔＰＡ外显子片段作探针，用地高辛标记进行菌落原位杂交，获得六个阳性菌落。提取阳性菌落质粒，经限制性内切酶Ｓａｌ Ⅰ和ＥｃｏＲⅠ酶切分析鉴定，得到３个ｔＰＡｃＤＮＡ重组子。
The human tissue - type plasminogen activator (tPA ) gene expresses at human earlystage embryo;; but the content of the tPA mRNA is very low. In this research we depuratedtie iPA mRNA from 17 days human embryo;; and founded cDNA library with the plas midPSTORT 1. The total number of the recombinants was 1.6 ×106. which could include thevery lower expression gene cDNA. A 17s bp tPAexon labeled with the digoxigeninas aprobe. We got six positive bacterial colonies by bacterinl original place hybridization;; and gotthree tPA cDNA recombinants by the identification with the restriction enzyme sall andEcoRI.
出处
《生物技术》
CAS
CSCD
1999年第1期1-3,共3页
Biotechnology
