CONSTRUCTION AND IDENTIFICATION OF HCMV cDNA EXPRESSING LIBRARY AND SCREENING OF pp65 POSITIVE CLONES
- VernacularTitle:人巨细胞病毒cDNA文库的构建、鉴定及pp65阳性克隆筛选
- Author:
Jun HAN
;
Yan-Qiu LI
;
Yong-Zhong JIANG
;
Li YU
;
Ming-Li WANG
;
- Publication Type:Journal Article
- Keywords:
Human cytomegalovirus, pp65,cDNA expressing library
- From:
Microbiology
1992;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
In order to provide effective tool for further studying of the function of HCMV genome, developing of molecular vaccine and diagnostic reagents. Extraction of HCMV mRNA from HF cell infected by HCMV AD169 strain for 96h was reverse transcripted into cDNA, then was cloned into EcoR I-digested lambda gt11. HCMV AD169 strain cDNA expressing library has been constructed after packaging. The volume and the recombination rate of the prime cDNA expressing libraries was 3.6?10 6 and 96%, 168 positive clones of HCMV were screened by immune blotting with anti-HCMV mouse convalescent sera, 34 positive clones were obtained by dot nucleic acid hybridization with DIG-labled HCMV pp65 gene probe. 2 positive clones were amplified by HCMV pp65 all length primer. The PCR product has been tested by southern-blotting.The PCR product was sequenced and was taken as homology comparison by DNASIS software,and the homology is 98%.To lay the foundation of furher cloning,expressing the pp65 gene,further studying of the function of the pp65 prodct.
