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植物乳杆菌melA基因的克隆及其作为食品级筛选标记的初步研究
引用本文:任大勇,李 昌,秦艳青,杜寿文,诸晴丽,金宁一. 植物乳杆菌melA基因的克隆及其作为食品级筛选标记的初步研究[J]. 食品科学, 2011, 32(9): 135-139. DOI: 10.7506/spkx1002-6630-201109031
作者姓名:任大勇  李 昌  秦艳青  杜寿文  诸晴丽  金宁一
作者单位:1.吉林农业大学食品科学与工程学院2.中国人民解放军军事医学科学院军事兽医研究所3.吉林大学畜牧兽医学院
基金项目:科技部重大传染病专项(2008ZX10004-015);军内“十一五”科技攻关项目(06G127);吉林省高新技术产业发展项目(2010);长春市科技特派员行动计划项目(09KT04)
摘    要:以L. plantarum1.557的基因组DNA为模板,通过PCR技术成功克隆α-半乳糖苷酶基因(melA基因),与报道的melA基因序列同源性达到99%以上。再以大肠杆菌-乳酸菌穿梭表达载体pMG36e为基本骨架,将melA基因插入该载体,构建melA基因标记的穿梭表达载体pMG36e-melA。重组表达质粒pMG36e-melA经Sac I和Sph I双酶切和PCR鉴定与预期片段大小相符。结果表明已初步构建了以melA基因为食品级筛选标记的重组载体。

关 键 词:α-半乳糖苷酶  melA  食品级筛选标记  穿梭表达载体  乳酸乳球菌  

Cloning of melA Gene from L. plantarum as a Food Grade Selection Marker
REN Da-yong,LI Chang,QIN Yan-qing,DU Shou-wen,ZHU Qing-li,JIN Ning-yi. Cloning of melA Gene from L. plantarum as a Food Grade Selection Marker[J]. Food Science, 2011, 32(9): 135-139. DOI: 10.7506/spkx1002-6630-201109031
Authors:REN Da-yong  LI Chang  QIN Yan-qing  DU Shou-wen  ZHU Qing-li  JIN Ning-yi
Affiliation:1. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China;2. Military Veterinary Institute, Academy of Military Medical Sciences of the PLA, Changchun 130062, China;3. College of Animal and Veterinary Medicine, Jilin University, Changchun 130062, China
Abstract:A full-length α-galactosidase gene (melA) sharing 99.2% identity with that previously reported melA (accession number: AF189765) was amplified by polymerase chain reaction (PCR) from L. plantarum 1.557 The PCR-amplified melA gene was cloned into the E. coli-L. plantarum shuttle vector pMG36e and transformed into E. coli DH5α. After the transformant was identified, the recombinant plasmid was purified and electrotransformed into L. lactis. The transformant containing melA gene was easy to detect by the observation of yellow colonies in BCP medium containing melibiose. This result indicated that α-galactosidase gene could be used as a maker for food grade selection.
Keywords:α-galactosidase  melA gene  food grade selection marker  shuttle expression vector  Lactococcus lactis  
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