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Titolo:
Expression of human beta-subunit nerve growth factor (hNGFB) in yeast Pichia pastoris and E-coli
Autore:
Fan, Q; Liu, HD; Sun, T; He, RQ;
Indirizzi:
Chinese Acad Sci, Inst Microbiol, Beijing 100080, Peoples R China Chinese Acad Sci Beijing Peoples R China 100080 100080, Peoples R China Chinese Acad Sci, Inst Biophys, Beijing 100101, Peoples R China Chinese Acad Sci Beijing Peoples R China 100101 100101, Peoples R China
Titolo Testata:
CHINESE SCIENCE BULLETIN
fascicolo: 6, volume: 44, anno: 1999,
pagine: 528 - 533
SICI:
1001-6538(199903)44:6<528:EOHBNG>2.0.ZU;2-X
Fonte:
ISI
Lingua:
ENG
Soggetto:
SACCHAROMYCES-CEREVISIAE; MOUSE; GENE; SEQUENCE;
Keywords:
hNGFB; P-pastoris; secretive expression vector; pET-15b; fusion protein;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Physical, Chemical & Earth Sciences
Citazioni:
15
Recensione:
Indirizzi per estratti:
Indirizzo: Fan, Q Chinese Acad Sci, Inst Microbiol, Beijing 100080, Peoples R China Chinese Acad Sci Beijing Peoples R China 100080 , Peoples R China
Citazione:
Q. Fan et al., "Expression of human beta-subunit nerve growth factor (hNGFB) in yeast Pichia pastoris and E-coli", CHIN SCI B, 44(6), 1999, pp. 528-533

Abstract

The gene hNGFB encoding the beta subunit of human nerve growth factor (hNGF) was cloned into P. pastoris secretive expression vector pHIL-S1 and E. coli expression vector pET-15b. The recombinant hNGFB vectors pSNGF and pET15b-NGF were transformed into P. pastoris host cell GS115 (Mut(+), His(-)) and E. coli Strain BL21 (DE3) respectively. Expression and secretion of hNGFB in P. pastoris was attempted under the direction of the AOX1 promoter andPHO1 signal sequence. The positive colonies growing on medium without histidine were further selected by PCR. The yield of rehNGFB in GS115 was about14.4% of total cellular secretive protein. The secreted protein was immunological active on Western blotting with rabbit anti-mNGFB antibodies. The fusion protein yield of rehNGFB in E. coli BL21 (DE3) was about 10.3% of total cellular protein after IPTG induction. Western blot detection showed itsimmunological activity.

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Documento generato il 22/09/20 alle ore 14:24:23