Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2011, Vol. 33 ›› Issue (6): 649-653 .doi: 10.3881/j.issn.1000-503X.2011.06.013

• Original Articles • Previous Articles     Next Articles

Adenovirus Construction of Expression and It's Function of Connective Tissue Growth Factor

GONG Ning, ZHANG Hua-bing, FANG Fu-de, CHANG Yong-sheng   

  1. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences,CAMS and PUMC, Beijing 100005, China
  • Received:2011-10-17 Revised:2012-01-04 Online:2011-12-20 Published:2011-12-20
  • Contact: FANG Fu-de
  • Supported by:

    Supported by the National Basic Research Program of China (973 Program) (2011CB504004)

Abstract: Objective To construct and identify a adenovirus vector of the expression of connective tissue growth factor(CTGF) and to explore the role of CTGF in the metabolism of glucose and lipid. Methods The over-expressed plasmid of CTGF was cloned, and then the CTGF sequences were cloned into pAdTrack-CMW vector. The reformed E.coli BJ5183-sensitive bacteria that contain pAdEasy-1 were transformed with lined vector cut by Pme I enzyme. The recombinant adenovirus vector was cut with Pac I enzyme and obtained, then transfected 293A cells to produce virus. Through three times of amplification, the adenovirus infected the primary hepatocyes to determine the infection efficiency and CTGF expression. The mice were starved for several time periods, and then the liver RNA was extracted for real-time PCR to detect the expressions of CTGF under different nutritional conditions. Results The adenovirus of CTGF was successfully produced with an infection efficiency of 90%. The expressions of the CTGF were different under different nutritional conditions and showed a coincidence with the expression of peroxisome proliferators-activated receptor γ coactivator 1α. After the mice were starved for 24h, the expression of CTGF increased by (2.38±0.51) folds; after the mice were starved for 48 h, the expression of CTGF increased by (2.95±0.57) folds (P<0.05). Conclusion CTGF is speculated to be involved in the metabolism of glucose and lipids.

Key words: connective tissue growth factor, over-expression, adenovirus, starving

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