Construction of cDNA Library From Human Nasopharyngeal Carcinoma Cell HNE_2
- VernacularTitle:鼻咽癌细胞HNE_2cDNA文库的构建
- Author:
Jun SHU
;
Guancheng LI
;
Xiaojuan HE
- Publication Type:Journal Article
- Keywords:
Nasopharyngeal carcinoma;
cDNA library;
Gene cloning
- From:
Journal of Chinese Physician
2001;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a cDNA library from human nasopharyngeal carcinoma cell HNE 2.Methods The total RNA was separated from human NPC(nasopharyngeal carcinoma)cell HNE 2 and the mRNA was isolated from the total RNA by MagneSphere technique,then the first-strand cDNA was synthesized with oligo(dT) primer containing sfiI site while the double-strand cDNA was amplified through LD-PCR(Long-distance PCR) by SMART technique.The double-strand cDNA was digested by sfiI(IA &IB)restriction enzyme before cDNA size fractionation ,the double-strand cDNA fractionated was ligated into the ?TripIEx2 vector and then was packaged in vitro.Results The unamplified human NPC cell HNE 2 cDNA library consists of 0 78?10 6 independent clones,and the percentage of recombinant clones was more than 96%.The titer of the amplified cDNA library was 1 02?10 9 pfu/ml and the average insert of the recombinants was 1 2kb.Conclusions The quality of the constructed human NPC cell HNE 2 cDNA library is excellent and helpful to screen NPC specific-antigen.
