Optimization of Prokaryotic Expression of Excretory-secretory Antigen Ts8B1 Protein Gene in Cysticercus cellulosae

doi:10.7668/hbnxb.2009.03.013

Abstract

Abstract: In order to obtain the highest expression level of excretory2secretory antigen Ts8B1 protein gene of Cys2 ticercus cellulosae in Escherichia coli,the factors affecting the protein expression were studied,including the medium,cul2 ture temperature,inducing time,the final concentration of inductor IPTG and Ampicillin (Amp),and the temperature change before and after induction. The results indicated that the expression of pGEX26P₂1/ Ts8B1 fusion protein reached the highest level when TB culture medium was used at 37 ℃for 3 h and then induction culture was carried out overnight at 32 ℃using 0. 2 mmol/L IPTGand 200 mmol/L Amp. The expression fusion protein accounted for 33 %of the total cell lysates. SDS2PAGE analysis showed that the molecular mass of pGEX26P₂1/ Ts8B1 fusion protein was about 33 kDa,the same as predicted. Western blot analysis indicated that the Cysticercus cellulose polyclonal antibody could be specifically bound to pGEX26P₂1/ Ts8B1 fusion protein. The results would be useful for obtaining the maximum yield of fusion protein and for researching its application value in detecting cysticercosis.

Key words: Cysticercus cellulosae, Excretory2secretory antigen, Ts8B1 protein, Prokaryotic expression

CLC Number:

WANG Qiu xia,, ZHANG Mei ying, ZHANG Gai ping, WANG Xuan nian,, NING Chang shen, LU Kun,, ZHANG Long xian, JIAN Fu chun. Optimization of Prokaryotic Expression of Excretory-secretory Antigen Ts8B1 Protein Gene in Cysticercus cellulosae[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(3): 59-63. doi: 10.7668/hbnxb.2009.03.013.

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