Construction of lymphocyte subtracted cDNA library of unstable angina
- VernacularTitle:构建不稳定型心绞痛淋巴细胞差异表达cDNA消减文库
- Author:
Bin ZHANG
;
Xueqing LIANG
;
Xiyong YU
;
Jianzhan FENG
- Publication Type:Journal Article
- Keywords:
Angina, unstable;
Suppression subtractive hybridization;
Subtractive cDNA library
- From:
Chinese Journal of Pathophysiology
1999;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct a subtracted cDNA library of differentially expressed genes in human unstable angina lymphocytes. METHODS: Suppression subtractive hybridizations (SSH) were performed between the patients with unstable angina pectoris and stable angina pectoris. Lymphocyte RNA, the obtained forward and reverse cDNA fragments were directly inserted into T/A cloning vector and transformed into E.coli JM 109 to construct a subtractive cDNA library. The inserting fragments were screened by blue and while blot screening and bacterium liqulid PCR. RESULTS: Each subtractive cDNA library contained more than 2000 positive bacteria clones. Most of them distributed between 200-600 bp inserts. CONCLUSION: The library is efficient and lays solid foundation for screening and cloning new and specific expressed genes in unstable angina lymphocyte RNA.