[Study on the cloning, in vitro expression and bioactivity of Ailuropoda melanoleuca Qinling subspecies interleukin-2].
OBJECTIVE
To identify if any difference exisit between the population of Sichuan and Qinling giant panda. express IL-2 protein in BL21(DE3)with biological activity.


METHODS
IL-2 was amplified by RT-PCR from adult Qinling giant panda peripheral blood lymphocyte which was induced by ConA, and cloned into prokaryotic expression vector of pET32a-IL-2. The fusion protein was expressed in BL21(DE3)/pET system and identified by SDS-PAGE and Western blot; inoculate rabbit with purified expression products to prepare polyclonal antibody; use lymphocytes proliferation in vitro to detect biological activity.


RESULTS
The protein of IL-2 was obtained by recombination expression, molecule weight is 34 000 .The specificity of polyclonal antibody was obtained, in vitro activity test indicated that the recombinant protein IL-2 having an activity of promoting the proliferation of lymphocytes. The effect can be stopped by polyclonal antibody which was prepared before.


CONCLUSIONS
The IL-2 gene of Qinling giant panda was cloned and expressed in E.coli successfully, and the homology of IL-2 gene in these two population is 99.4% and the recombination protein can promote the proliferation of lymphocytes in vitro from giant panda.
Qiao
Qi
Q
College of Veterinary Medcine, Northwest A & F University, Yangling 712100, China.
Ma
Qing-yi
QY
Zhu
Hui
H
Chen
De-kun
DK
D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002681:China; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D007376:Interleukin-2 / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D010802:Phylogeny; D011994:Recombinant Proteins / Q000378:metabolism* / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D001503:Ursidae / Q000145:classification / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D007376:Interleukin-2; D011994:Recombinant Proteins