Journal of Capital Medical University ›› 2012, Vol. 33 ›› Issue (2): 187-192.doi: 10.3969/j.issn.1006-7795.2012.02.011

• 基础研究 • Previous Articles     Next Articles

Establishment of stably transfected cell line expressing BβArg448 or BβLys448 fibrinogen

LIN Yuan1,2, LI Ji-feng1,2, LIU Jie1,2, NIU Meng-lin2, SUN Ran2, WANG Jun1,2, LIU Yan1,4, WANG Chen1,3   

  1. 1. Department of Respiratory and Critical Care, Beijing Chaoyang Hospital, Capital Medical University, Beijing Key Laboratory of Respiratory and Pulmonary Circulation, Beijing Institute of Respiratory Medicine, Beijing 100020, China;2. Department of Physiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;3. Beijing Hospital, Ministry of Health, Beijing 100730, China;4. Department of Cardiac Surgery, Being Chaoyang Hospital, Capital Medical University, Beijing 100020, China
  • Received:2011-12-26 Revised:1900-01-01 Online:2012-04-21 Published:2012-04-21

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Abstract: Objective To investigate the effect of BβArg448Lys polymorphism on the function of fibrinogen, and to establish the stably transfected cell line expressing fibrinogen with BβArg448 and BβLys448 polymorphism. Methods Mutant fibrinogen Bβ chain cDNA(BβLys448) pMLP-FGB(448A) was generated from wild type Bβ chain cDNA(BβArg448) pMLP-FGB(448G), which contains the entire fibrinogen Bβ chain cDNA using site-directed mutagenesis method. The expression vectors encoding the entire fibrinogen Aα chain, wild type/ mutant type Bβ chain and γ chain cDNA were introduced into CHO-K1 cells respectively using Lipofectamine. The expression of all three chains in the transfected cell line was tested by RT-PCR. Results The stably transfected cell line expressing fibrinogen with BβArg448 and BβLys448 polymorphism was successfully established, which provide a useful cell model to the further functional study.Conclusion The stably transfected cell line expressing fibrinogen with BβArg448Lys and BβLys448 polymorphism was successfully established, which provide a useful cell model to the further functional study.

Key words: fibrinogen, site-directed mutagenesis, stable transfected cell line

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