重组猪表皮生长因子在酿酒酵母中的克隆表达及其生物学活性鉴定

doi:10.11843/j.issn.0366-6964.2014.12.009

Abstract

Abstract:

Porcine epidermal growth factor (EGF) can induce effects on DNA synthesis and stimulation of cellular proliferation in intestine to improve the performance，gut development and immune function of early weaned-piglets.Herein，generation of Neijiang pig EGF-expressing Saccharomyces cerevisiae (S.cerevisiae) was beneficial to livestock production and clinical research；pYES2-Mfα-spEGF were originated from synthetic optimized EGF from breast tissue of Neijaing pig (pEGF) and signal peptide (Mfα) fragments of S.cerevisiae were cloned into pYES2/CT.Recombinant S.cerevisiae INVSc1(pYES2-Mfα-spEGF) expressing EGF protein was generated by transforming the plasmid pYES2-Mfα-spEGF into S.cerevisiae INVSc1.The target EGF protein was determined by Tricine-SDS-PAGE electrophoresis and Western blot，meanwhile，its biological activity was also determined in vivo and in vitro.The spEGF-expressing recombinant S.cerevisiae INVSc1(pYES2-Mfα-spEGF) was constructed successfully and expressed in S.cerevisiae.The target protein EGF was identified by Tricine-SDS-PAGE electrophoresis and Western blot.The results of biological activity detection showed that spEGF-expressing protein could stimulate enterocyte proliferation (P<0.05)，improve the growth performance (e.g.，body weight，average daily gain，feed intake，and feed conversion rate) (P<0.05)，gut development (e.g.，villus height，crypt depth，and contents of total protein，DNA，and RNA) (P<0.05)，and immune function (e.g.，IgA，IgG，and IgM) of early-weaned SD rat (P<0.05).The concentration of target protein from the supernatant fluid of INVSc1(pYES2-Mfα-spEGF) is about 30 mg•L^-1，and INVSc1(pYES2-Mfα-spEGF) have some prominent features encompassing biological activity to be directly applied to livestock production and clinical research.

CLC Number:

S828
S815.4

WANG Shu-jin，ZHOU Lin，CHEN Hui-na，ZHANG Zheng-fan，HUANG Yan-ling，GUO Chun-hua,WANG Yong. Cloning and Expression of Recombinant Pig Epidermal Growth Factor in Saccharomyces cerevisiae and Its Biological Activity[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(12): 1971-1980.

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Cloning and Expression of Recombinant Pig Epidermal Growth Factor in Saccharomyces cerevisiae and Its Biological Activity

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