[Expression of Crg-2 by a bacteria recombinant system of adenovirus].
OBJECTIVE
To express an immunochemotactic and angiostatic anti-tumor molecule, CXC chemokine Crg-2, by E. coli recombinant system of adenovirus.


METHODS
By E. coli recombination system of adenovirus, the shuttle vector pShuttle-cmv/crg-2 was co-transfected into BJ5183 E. coli with adenovirus backbone plasmid pAdEasy-1. The recombinants were transfected into 293 packaging cells and adenovirus was packaged and amplified. The molecular weight and bioactivity of recombinant protein were determined by Western blot and chemotaxis assay, respectively.


RESULTS
Recombinant adenovirus genome backbone bearing crg-2 gene, pAd/crg-2, was successfully obtained as evidenced by endonucleases digestion and PCR. Adenovirus was packaged and amplified to a titre of 4x10(9) TCID50/L. The cultured supernatant of the infected 293 cells contented a protein of nearly 10,000 and presented a significantly chemotatic effect towards activated spleen lymphoblasts.


CONCLUSIONS
Recombinant adenovirus Ad/crg-2 obtained by E. coli recombinant system of adenovirus can efficiently express bioactive Crg-2 chemokine.
Ning
Han-bing
HB
State Key Laboratory of Cancer Biology, Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Zhao
Li-na
LN
Li
Ting-ting
TT
Wang
Xiao-xia
XX
Li
Ji-chang
JC
Liu
Zhi-guo
ZG
Fan
Dai-ming
DM
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D054357:Chemokine CXCL10; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D015846:Monokines / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
D054357:Chemokine CXCL10; C485433:Cxcl10 protein, mouse; D015846:Monokines; D011994:Recombinant Proteins