EFFICIENT SCREENING OF RETROVIRAL CDNA EXPRESSION LIBRARIES

被引：224

作者：

KITAMURA, T ^{[1
]}

ONISHI, M ^{[1
]}

KINOSHITA, S ^{[1
]}

SHIBUYA, A ^{[1
]}

MIYAJIMA, A ^{[1
]}

NOLAN, GP ^{[1
]}

机构：

[1] STANFORD UNIV,DEPT MOLEC PHARMACOL,STANFORD,CA 94305

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1995年 / 92卷 / 20期

关键词：

D O I：

10.1073/pnas.92.20.9146

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Expression cloning of cDNAs was first described a decade ago and was based on transient expression of cDNA libraries in COS cells. In contrast to transient transfection of plasmids, retroviral gene transfer delivers genes stably into a wide range of target cells. We utilize a simple packaging system for production of high-titer retrovirus stock from cDNA libraries to establish a cDNA expression cloning system. In two model experiments, murine interleukin (IL)-3-dependent Ba/F3 cells were infected with libraries of retrovirally expressed cDNA derived from human T-cell mRNA or human IL-3-dependent TF-1 cell line mRNA, These infected Ba/F3 cells were selected for the expression of CD2 by flow cytometry or for the alpha subunit of the human IL-3 receptor (hIL-SR alpha) by factor-dependent growth, CD2 (frequency, 1 in 10(4)) and hIL-3R alpha (frequency, 1 in 1.5 x 10(5)) cDNAs were readily detected in small-scale experiments, indicating this retroviral expression cloning system is efficient enough to clone low-abundance cDNAs by their expression or function.

引用

页码：9146 / 9150

页数：5

共 22 条

[1] EFFICIENT RETROVIRAL-MEDIATED GENE-TRANSFER INTO HUMAN B-LYMPHOBLASTOID CELLS EXPRESSING MOUSE ECOTROPIC VIRAL RECEPTOR [J].