ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2005, Vol. 36 ›› Issue (9): 877-881.doi:

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The Eukaryotic Expression of Porcine LH-β Gene in vitro and the Effect of Transgenetic Expression in vivo on the Reproduction of Mice

L Xue-bin; LI Jiang-ling;GAO Rong; WU Mei;YANG Yi;WANG Li-huan; LIU Shi-gui   

  1. 1.Sichuan Academy of Animal Science, Chengdu 610066, China;2.College of Life Science, Sichuan University, Chengdu 610064,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-09-25 Published:2005-09-25

Abstract: VR1020 was first used as the eukaryotic expression plasmid to construct LH-β eukaryotic expression system. The recombinant plasmid was detected by the plasmid PCR and BamHⅠ,BglⅡ digestion to identify the correct insert orientation, the new plasmid was named VPLH-β. VPLH-β was transformed into E. coli DH5α competent cells. The ability of VPLH-β to express full-length porcine LH-β in vitro was confirmed by RT-PCR in Cos7 cells that were transiently transfected with VPLH-β in the entrapment of Lipofectamine. 48 BALB/c female mice at the age of 2 weeks were divided into 3 groups and were vaccinated intramuscularly in the left and right quardriceps respectively with 100μg VPLH-β, with 100μg VR1020; Saline solution was used as control treatment. The blood of mice were collected from tail vain on 7, 14, 21, 28 days after inoculation to determine the dynamic changes of the content of LH-β and FSH in the sera. The reproduction of mice was also detected. The experimental results indicated that the VPLH-β, we first recombined, could correctly express in vitro by using the Lipofectamine entrapment system. In the animal research, the amount of LH and FSH were significantly raised in the mice inoculated with VPLH-β plasmid, which was correlated with reproduction performance of the mice that produced more young mice than the control mice. These results indicate that the inoculation with VPLH-β could remarkably raise the content of LH and FSH in the sera of mice and promote the reproduction performance.