%0 Journal Article
%T Construction of a Flp "Exchange Cassette" Contained Vector and Gene Targeting in Mouse ES Cell<br>含FLP“交换盒”结构的打靶载体构建及ES细胞打靶研究
%A J M Zheng
%A J Li
%A J L Fu
%A <br>郑敬民
%A 李坚
%A 傅继梁
%J 生物工程学报
%D 2001
%I 
%X Using the HPRT genomic DNA fragment and synthesized oligonucliotides, pSP-HPRT-F-Neo-F3 was designed and constructed as a replacement gene targeting vector by usual molecular cloning techniques. Structure of pSP-HPRT-F-Neo-F3 was identified by restrictive digestion analysis and partly sequencing. Then linearized pSP-HPRT-F-Neo-F3 DNA was electroporated into ES cells, and transfected cells were screened by being cultured in medium containing 200 micrograms/mL G418 and 2 micrograms/mL 6-GT. Twenty-four double drug resistant clones were picked up and analyzed, among them, two clones were proved to have taken place the required recombination by PCR and southern blotting analysis.
%K gene targeting
%K ES cell
%K HPRT
%K FLP recombinase<br>基因打靶
%K 胚胎干细胞
%K HPRT
%K FLP重组酶
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=41186F835F85CA7A&yid=14E7EF987E4155E6&vid=BCA2697F357F2001&iid=94C357A881DFC066&sid=CDC418F38C4BFD60&eid=1FA4E9C3E6E88FC8&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=8