Transformation of, and Heterologous Protein Expression in, Lactobacillus agilis and Lactobacillus vaginalis Isolates from the Chicken Gastrointestinal Tract
Transformation of, and Heterologous Protein Expression in, Lactobacillus agilis and Lactobacillus vaginalis Isolates from the Chicken Gastrointestinal Tract
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鸡胃肠道分离乳杆菌和阴道乳杆菌的转化和异源蛋白表达
DOI：
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发表时间：
2010
影响因子：
4.4
通讯作者：
G. Allison
中科院分区：
文献类型：
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作者：
David P. Stephenson;R. Moore;G. Allison
摘要乳杆菌在鸡的胃肠道中是天然存在的，利用原生菌株运送治疗蛋白很有兴趣。此前，我们鉴定了三株鸡源乳杆菌，Agilis乳杆菌La3，阴道乳杆菌Lv5和卷曲乳杆菌Lc9，它们持续存在于商业饲料或高蛋白饲料喂养的鸡的胃肠道中。在目前的研究中，我们研究了这些菌株的电转化能力，确定了质粒载体的稳定性，并比较了几种不同启动子引导的报告基因的表达。La3和Lv5菌株的转化效率分别为每微克DNA 108个和106个转化子。测试的第三个菌株，卷曲乳杆菌Lc9，对所研究的所有转化方案都是顽抗的。PTRK563和pTRKH2分别在La3和Lv5中保持了100代以上。用异源启动子和同源启动子分析了La3和Lv5表达外源报告基因GFP的能力。含有La3ldhL启动子的La3和Lv5转化子的荧光强度最高。据我们所知，这是第一个成功转化并在阴道乳杆菌中表达异源蛋白的报道。这些菌株在体外表达异源蛋白的能力表明它们作为治疗性多肽的体内递送载体到鸡的胃肠道具有潜在的用途。
ABSTRACT Lactobacilli are naturally found in the gastrointestinal tract of chickens, and there is interest in utilizing autochthonous strains for the delivery of therapeutic proteins. Previously we identified three chicken-derived Lactobacillus strains, Lactobacillus agilis La3, Lactobacillus vaginalis Lv5, and Lactobacillus crispatus Lc9, which persist in the gastrointestinal tract of chickens fed either a commercial or high-protein diet. In the current study, we investigated the ability to electrotransform these strains, determined plasmid vector stability, and compared reporter gene expression directed by several different promoters. The La3 and Lv5 strains were reproducibly transformed with efficiencies of 108 and 106 transformants per microgram of plasmid DNA, respectively. The third strain tested, L. crispatus Lc9, was recalcitrant to all transformation protocols examined. The plasmid vectors pTRK563 and pTRKH2 were maintained over 100 generations in La3 and Lv5, respectively. The ability of La3 and Lv5 to express the heterologous reporter gene gfp was analyzed using heterologous and homologous promoters. Transformants of both La3 and Lv5 containing the La3 ldhL promoter were the most fluorescent. To our knowledge, this is the first report of successful transformation and heterologous protein expression in L. agilis and L. vaginalis. The ability of these strains to express heterologous proteins in vitro indicates their potential utility as in vivo delivery vectors for therapeutic peptides to the chicken gastrointestinal tract.
从鼠胃肠道分离的乳杆菌菌株中的隐性质粒。
影响因子：
4.4
作者：
Lin,JH;Savage,DC
通讯作者：
Savage,DC