title: [Cloning, expression of human keratinocyte growth factor and its purification and identification].
abstract: To clone KGF-2 gene, get hKGF-2 protein and detemine its activity. The cNDA of human KGF-2 was isolated from fetal lung by RT-PCR and cloned into pBV220 plasmid. The recombinant pBV220-hKGF-2 plasmid was transformed into E. coli (BL21), induced at 42 degrees C for the expression of hKGF-2. Recombinant human KGF-2 was purified from the ultrasonic-treated BL21 by heparin-Sepharose CL-6B treated column chromatography and cation exchange column chromatography. MTT method was used for the determination of its biological activity. SDS-PAGE showed that rhKGF-2 was expressed in E. coli BL21 as soluble protein of approximately 20kD. The rhKGF-2 protein can stimulate the proliferation of NIH3T3 cells significantly from 1 ng/mL to 10 ng/mL. HKGF-2 cDNA wasclned and highly expressed in E. coli BL21 and the purified rhKGF-2 showed the mitogenic activity on NIH3T3 cells.
authors.affiliation: National Laboratory of Molecular Virology and Genetic Engineering, Institute of Virus Prevention and Control, Chinese Center of Disease Control, Beijing 100052, China.
authors.forename: Bin-Wen
authors.initials: BW
authors.lastname: Wu
authors.forename: Zhao-Jun
authors.initials: ZJ
authors.lastname: Duan
authors.forename: Wu-Ping
authors.initials: WP
authors.lastname: Li
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Hong-Liang
authors.initials: HL
authors.lastname: Lü
authors.forename: Zuo-An
authors.initials: ZA
authors.lastname: Yi
authors.forename: Cheng-Hai
authors.initials: CH
authors.lastname: Zhang
authors.forename: Ju-Sheng
authors.initials: JS
authors.lastname: Lin
authors.forename: Jia-Long
authors.initials: JL
authors.lastname: Wang
authors.forename: Yun-De
authors.initials: YD
authors.lastname: Hou
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005333:Fetus; D051526:Fibroblast Growth Factor 10 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008168:Lung / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D051526:Fibroblast Growth Factor 10; D011994:Recombinant Proteins