Journal of South China University of Technology (Natural Science Edition) ›› 2005, Vol. 33 ›› Issue (1): 88-91,98.

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Expression and Modification of the M utant of Human Acidic Fibroblast Growth Factor

Zheng Qing1  Su Zhi-jian1  Wu Xiao-ping2  Xu Hua2  Huang Zhi-feng1  Zhao Wen1  Yao Cheng-can2  Huang Ya-dong1  Li Xiao-kun2   

  1. 1.Biopharmaeeutieal Research& Development Center,Jinan Univ.,Guangzhou 5 10632,Guangdong,China;2.Key Laboratory of Biopharmaceutics of Guangdong Province,Jinan Univ.,Guangzhou 510632,Guangdong,China
  • Received:2004-01-07 Online:2005-01-25 Published:2005-01-25
  • Contact: 郑青(1964-)，男，博士，副教授，主要从事生物制药方面的研究 E-mail:qz01cn@yahoo.com.cn
  • About author:郑青(1964-)，男，博士，副教授，主要从事生物制药方面的研究
  • Supported by:

    广东省自然科学基金资助项目(010424)；863计划资助项目(2002AA2Z3318)

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Abstract:

The codons of the 98 th and the 1 32nd cysteine(Cys)of haFGF(Human Acidic Fibroblast Growth Fac-tor)cDNA Were reFIlaced with the serine(Ser)codon by means of PCR.Thus,the recombinant plasmid pET3c-haFGFSer98,132 was constructed,which was then transformed into E coli BI21(DE3),with an expression level of 23.48%.The activity of the product was detected by MTT method,with the conclusion that the mutant of haFGFSer98,132 is of an activitv similar to stan dard haFGF. Maleimide-6-aminocaproyl-mPEG5000 ester was site-directed couple to the 31 st cysteine residue of haFGFSer98,132 .The result indicates that the modification rate is up to 30% ,and the re-mained specific activity of the purified PEG-haFGFSer98,132 is 55.53% .

Key words: human acidic fibroblast growth factor, site-directed mutation, mutant