Search colorectal cancer antigen genes by screening the colorectal tissue cDNA phage expression libraries with SEREX
- VernacularTitle:从大肠癌组织cDNA表达文库筛选肿瘤抗原基因
- Author:
Jingdong HE
;
Gaosu ZHOU
;
Bing XIAO
- Publication Type:Journal Article
- Keywords:
colorectal neoplasms;
SEREX (serological identification of antigens by recombinant expression cloning);
antigenic gene;
cDNA expression library
- From:
Medical Journal of Chinese People's Liberation Army
1983;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective Identification of colorectal cancer specific antigens from cDNA phage expression library by recombinant expression cloning (SEREX). Methods The cDNA phage expression library derived from colorectal cancer tissue was constructed using the SMART (Switching Mechanism at 5'end of RNA Transcript) techniques. The cDNA phage expression library was screened with SEREX (serological identification of antigen by recombinant cDNA expression libraries), the positive clones encoding antigenic genes were obtained after immunoscreening, and the nucleotide sequences of cDNA inserts were determined and analyzed with BLST software in GenBank, and the antigenic gene function was analyzed by bioinformatics. Results The cDNA phage expression library derived from colorectal cancer tissues was successfully constructed. The primary library consisted of 2.39?10 6 recombinants, and the recombinant rate was more than 97.5%. The titer of the amplified cDNA phage expression library was 4.1?10 10pfu/ml, and the size of inserted cDNA varied from 0.5 to 4.0kb. Sixteen positive clones encoding antigenic genes were obtained after immunoscreening, and results showed that 16 reactive clones were derived from 12 different genes. Ten of 12 genes were highly homologous to the genes known in GenBank, such as IFITM1, CD24, Survivin, KLK6, et al. Two expressed sequence tags (ESTs) were not found in GenBank. The antigenic genes included structural gene, regulation gene and metabolizing gene. Conclusion The tumor-associated antigen genes and the two ESTs were obtained by SEREX were worthy of further study on their structures and functions.
