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Published on: September 25, 2011
[Construction and identification of cDNA library for colorectal carcinoma antigens]
1Department of Cell Biology, China Medical University, Shenyang, China. Infsch@hotmail.com
Aim:
To construct a cDNA phage expression library of colorectal carcinoma antigens for screening colorectal carcinoma specific-antigens.
Methods:
The total RNA was extracted from two colorectal carcinoma cell lines CCL-187 and CX-1 . The mRNA from total RNA was isolated to synthesize the first and second strand cDNA. The ds-cDNA termini were blunted with pfu DNA polymerase. The blunted cDNAs were ligated with EcoR I adapters and then digested by Xho I. Small cDNA molecules (<400 bp) were removed through size fraction. After the cDNAs were inserted into ZAP expression vector, the ligated products were packaged in vitro and the bacteriophage particles infected the host strains XL1-Blue MRF'.
Results:
The efficiency of the primary was 2.3x10(9) pfu/L, while the recombination rate reached to 97%. The average length of the inserted fragment was over 1 kb.
Conclusion:
The quality of colorectal carcinoma cDNA phage expression library is excellent and helpful to screen colorectal carcinoma specific-antigens.
