ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (5): 670-675.doi:

• 遗传繁育 • Previous Articles     Next Articles

Recombination of CBH Ⅱ Gene with Non-antibiotic Selected Vector and Expression and Activity Detection in Lactobacillus

ZHAO Ying1,2，SUN Zhe1*， LIU Yan3，HU Hou-ying4，CHEN Da-fang5，LOU Yu-jie2，ZHANG Hong-fu1*   

  1. 1.State Key Lab of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China; 3. College of Animal Science and Technology, Inner Mongolia Agricultural University, Huhhot 010018, China; 4. Institute of Animal Science and Veterinarian of Jinan, Jinan 250002, China; 5. Guizhou Province Livestock and Poultry Seedstock Breeding, Guiyang 550018,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

PDF (PC)

732

Abstract: Recombinant plasmids pMD18-T-CBHⅡ and pW425t, prokaryotic expression shuttle vector between E.coli and Lactobacillus, were digested with SmaI and SphI enzymes respectively. The purified CBHⅡ gene was subcloned into the expression vector pW425t. Thus,the recombinant pW425t-CBHⅡ was constructed, and then was transformed into the competence thyA genemutant Lactobacillus DOMLaS107. Treated lysates of bacterium were loaded directly on SDS-PAGE, and approximately 49.6 kD protein was observed, and recombinant Lactobacillus can produced clear hydrolysis halos on the Congo-Red-CMC plate.