转内切葡聚糖酶基因罗伊乳酸杆菌的构建及其肉鸡饲喂效果验证

doi:10.11843/j.issn.0366-6964.2015.05.013

Abstract

Abstract:

The objective of the study was to construct the transgenic Lactobacillus reuteri with the ability of exocellular secreting endoglucanase and hydrolyzing β-glucan and evaluate its feeding effect on broilers，which would establish the foundation for application of the transgenic L.reuteri in animal production.pLEM4157（cel） vector was constructed by PCR and restriction enzyme，then transformed into competent cells of L.reuteri XC1 for SDS-PAGE analysis，determination of endoglucanase activity and evaluation of feeding effect on broilers.The results of PCR amplification and sequence blast showed that endoglucanase gene celW was about 1 500 bp，the similarity of gene and amino acid sequence were 99% and 100%，respectively.Native-PAGE amd SDS-PAGE analysis showed that the molecular mass of celW gene mature peptide was approximately 52 ku，which was consistent to the predicted molecular weight.The endoglucanase activity in the extracellular and intracellular fraction in the transformed L.reuteri culture were（0.96±0.08） and（0.37±0.09） U•mL^-1，respectively.L.reuteri pLEM4157（cel） had not significant effect on growth performance of broilers，but could decrease pH in duodenum and jejunum at 21 d，promote the growth development of kidney，cloacal bursa，thymus and pancreatic at 21 d，increase the thymus and pancreatic index at 42 d，improve the A/G and the content of Ca and P at 21 d，and the content of glucose at 42 d.L.reuteri pLEM4157（cel） successfully secreted and expressed endoglucanase，displayed the ability to hydrolyze carboxymethyl cellulose and feeding effect on broier.

CLC Number:

S831
S815.5

WANG Lei,CAO Ping-hua,CAI Bei,GAO Yin,GAO Kang,YANG Yu-xin,YANG Ming-ming,YUAN Chao,CHEN Yu-lin. Construction of Transgenic Lactobacillus reuteri with Endoglucanase and Evaluation of Its Feeding Effectiveness on Broilers[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(5): 774-783.

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Construction of Transgenic Lactobacillus reuteri with Endoglucanase and Evaluation of Its Feeding Effectiveness on Broilers

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