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[Construction of pcDNA-BLC and expression of BLC in tumor cells]
Chun-hua Zou1, Ling Tian, Yu-quan Wei
1Key Laboratory of Biotherapy of Human Diseases, Ministry of Education, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To establish a stable transformants of B-lymphocyte chemoattractant (BLC) with the eukaryotic expression vector for investigation of its anti-tumor function further.
Methods:
The BLC full-length cDNA was amplified by RT-PCR from the total RNA isolated from the spleen of mouse. After sequence determination, the BLC full-length cDNA was cloned into the eukaryote expression vector pcDNA3.1 (+), the resulted recombinant was designated as pcDNA-BLC. Then pcDNA-BLC was transfected into murine tumor cell line colon 26 by LIPOFECTIN. The stable transformants were selected by G418 and identified by immunoblot assay. The bioactivity of BLC was confirmed by chemotaxis assay.
Results:
The recombinant pcDNA-BLC was successfully constructed. The stable transformants of BLC can express the bioactive BLC in tumor cell lines.
Conclusion:
The stable transformants can stably express BLC in tumor cell lines. These stable transformants will be useful for further study of the BLC biological effect on tumor and for the development of cancer vaccine.
