一种具杀菌功能的haFGF融合基因在毕氏酵母中的表达.pdf

( ) 2010 50 12 J urnal f Nanchang University( Medical Scien ce) 2010, V l. 50 N . 12 53 haFGF , , , , , , , ( 广东医学院生物学教研室, 广东湛江524023) : , DNA cecr pin AD haFGF 5, CADA F , pPICZA , X-33 SDS PA GE W estern bl t , CADAF , M TT , haFGF , : ; ; ; : R392. 11 : A : 1000- 2294( 20 10) 12- 0053- 05 The Fusion Expression of a Antimicrobial Peptide haFGF in Pichia Pastoris Cell PANG Sh-i feng FU Hong- i, ZHENGKe- in, LIAO Xia, LIANGPeng, CAO Ding-guo, CHEN Xiao-ping, ZHOU Ru-bing ( D ep ar tment of B iol og y , Guang dong M ed ic al Coll eg e, Zhanj iang 524023, Ch i na) ABSTRACT: Objective C nstructed a dual functi n peptide, and made it expressed in y east. Methods Cecr pin AD w as c nnected w ith 5end f the cDNA f human acidic Fibr blast Gr w th Fact r( haFGF) by using DNA r ec mbinati n in vitr , and then CADAF fusi n gene w as cl ned int pPICZA resulting in a secreti n expressi n vect r. T he rec mbinant plasmid w as tr ansf rmed int Pichia past ris X-33, w hich w as induced by methan l inducti n. Results SDS- PAGE and Wester n bl t analysis sh w ed the fusi n gene CADAF w as expr essed in Pichia past ris successfully, It was c nfirmed that CADAF pr tein has the immun genicity f haFGF. The analy- sis f restraining the gr w th f E c liK 12D31 sh w ed CADAF has the antimicr bial activity. MT T results indicated CADAF can stimulate Balb/ c3T 3 cells pr liferati n. Conclusion aFGF- Cecr pin fusi n peptide have been expressed in yeast successfully. This study pr vides a f unda- ti n f r the use f multifuncti nin fact r in w und cur ed. KEY WORDS: human acidic fibr blast gr w th fact r( haFGF) ; cecr pin; fusi n gene; pichia past ris ,