Expression of porcine epidermal growth factor in Pichia pastoris and its biology activity in early-weaned piglets.
Early-weaned piglets often have abnormalities in intestinal morphology and function. Epidermal growth factor (EGF) is critical in the development and in the repair of the gastrointestinal tract in pigs. This study investigated the effects of dietary EGF supplementation on growth performance and small intestinal morphology of early-weaned piglets. The functional domain of porcine EGF (pEGF) was cloned after RT-PCR amplification. The recombinant protein was expression by the Pichia pastoris expression system and the construct pPIC9K-pEGF was transformed into host GS115. The secretary recombinant protein in the supernatants was analyzed by SDS-PAGE. The gel indicated that the extra band at 6 kDa in the transformant, which corresponds to the standard hEGF, were both reactive to anti-pEGF antibody by Western blotting. The expression level of pEGF in the culture supernatant was 870 microg/mL. An animal feeding test was conducted to identify the effects of pEGF supplementation on growth performance and the development of digestive tracts of 14-day weaned piglets. The dietary treatment was a corn-soybean meal basal diet either with or without 1.5 mg/kg recombinant pEGF from the transformant fermentative supernatant. Dietary treatments enhanced the daily gain during 0-7 days postweaning (p < 0.05), but did not affect the performance throughout the entire test period. Dietary supplemental pEGF significantly increased serum IgA levels on day 18 postweaning, and increased the mucosa IgA levels and crypt depth at jejunum on day 28 postweaning (p < 0.05). The experimental results showed that the recombinant pEGF could be secreted by P. pastoris. The trophic effects of pEGF on growth performance, immune response, and small intestine development were determined by feeding recombinant pEGF to early-weaned piglets.
Lee
Der-Nan
DN
Department of Animal Science, National Ilan University, Ilan, Taiwan, ROC. dnlee@niu.edu.tw
Kuo
Tsun-Yung
TY
Chen
Ming-Cheng
MC
Tang
Tsung-Yin
TY
Liu
Fu-Hwa
FH
Weng
Ching-Feng
CF
D000818:Animals; D004815:Epidermal Growth Factor / Q000096:biosynthesis / Q000494:pharmacology*; D005260:Female; D006128:Growth / Q000187:drug effects; D007070:Immunoglobulin A / Q000096:biosynthesis; D007422:Intestines / Q000187:drug effects; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000494:pharmacology; D013552:Swine; D014886:Weaning
D007070:Immunoglobulin A; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor