[Construction of pcDNA-BLC and expression of BLC in tumor cells].
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.


CONCLUSIONS
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.
Zou
Chun-hua
CH
Key Laboratory of Biotherapy of Human Diseases, Ministry of Education, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Tian
Ling
L
Wei
Yu-quan
YQ
Zhao
Xia
X
Jiang
Yu
Y
Yi
Tao
T
Li
Qiu
Q
Li
Hong-li
HL
Tan
Guang-hong
GH
Deng
Hong-xin
HX
Li
Jiong
J
Wen
Yan-jun
YJ
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology; D000818:Animals; D019496:Cancer Vaccines / Q000096:biosynthesis; D054382:Chemokine CXCL13; D019743:Chemokines, CXC / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D018076:DNA, Complementary / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014407:Tumor Cells, Cultured
D019496:Cancer Vaccines; D054382:Chemokine CXCL13; D019743:Chemokines, CXC; C486386:Cxcl13 protein, mouse; D018076:DNA, Complementary; D011994:Recombinant Proteins