Isolation of cDNA Clones Using PAC and BAC Probes
- VernacularTitle:以PAC和BAC克隆为探针筛选cDNA文库
- Author:
Liwei HAN
;
Wenxin QIN
;
Xintai ZHAO
;
Yi HUANG
;
Pingping ZHANG
;
Dafang WAN
;
Jianren GU
- Publication Type:Journal Article
- Keywords:
bacteriophage P1-derived artificial chromosome;
bacterial artificial chromosome
- From:
Fudan University Journal of Medical Sciences
2000;27(6):449-452
- CountryChina
- Language:Chinese
-
Abstract:
Purpose To introduce a method to isolate cDNA clones using bacteriophage P1-derived artificial chromosome (PAC) or bacterial artificial chromosome (BAC) as probe for hybridization and try to find some novel genes related to hepatocellular carcinoma. Methods PAC 579 (D17S926 locus) and BAC 1529 (D17S1529 locus) in the deletion region of chromosome 17p13.3 in human hepatocellular carcinoma were chosen to screening the human liver cDNA library as probe for hybridization. The isolated positive cDNA clones were partially sequenced, then analyzed by computer comparison and Southern blot. Results After three cycles of screening, 78 and 8 candidate positive cDNA clones were isolated using PAC 579 and BAC 1529 probes respectively. Further analysis indicated 18 cDNA clones isolated by PAC 579 probe and 5 cDNA isolated by BAC 1529 probe were potential novel genes related to hepatocellular carcinoma. Conclusions The isolation of cDNA clones using PAC and BAC probes is effective and practical.
