%0 Journal Article
%T Green fluorescent protein labeling of ylyA gene in Bacillus subtilis<br>枯草杆菌ylyA基因的绿色荧光蛋白标记
%A Nairui Huo
%A Peter J Lewis
%A <br>霍乃蕊
%A Peter J. Lewis
%J 微生物学报
%D 2008
%I 
%X Objective] In this study we labeled the ylyA gene of Bacillus subtilis with green fluorescent protein (GFP) to investigate the subcellular localization of YlyA protein. Methods] Chromosomal DNA was prepared from different strains of Bacillus subtilis, ylyA amplified by PCR and the products sequenced. Full-length ylyA was then amplified and inserted into the GFP plasmid vector pSG1729, to give pNG426 containing a gfpmut1-ylyA fusion. Finally, Bacillus subtilis 168 was transformed with pNG426, resulting in insertion of the gfpmut1-ylyA fusion into the chromosome at the amyE locus. Double crossover integrants (subsequently named BS363) were identified by their inability to hydrolyse starch and verified by colony-PCR. Following induction of gfpmut1-ylyA expression with 0.5% xylose, localization of the fusion protein was determined by epifluorescence microscopy. Results] The correct sequence and translation start site of ylyA was identified from sequence analysis of the several amplified PCR products permitting construction of a gfpmut1-ylyA fusion. Microscopic observation of strain BS363 showed that the GFP labeled YlyA was distributed around the cell periphery, closely juxtaposed with the cytoplasmic membrane. Conclusion] GFP labeled YlyA produced by BS363 cultured on the nutrient agar solid medium distributed around the cell periphery.This suggests it may play a role in membrane biology.
%K Bacillus subtilis
%K YlyA protein
%K fluorescent labeling<br>枯草杆菌
%K YlyA蛋白
%K 荧光标记
%K 枯草杆菌
%K 基因
%K 绿色
%K 荧光蛋白
%K Bacillus
%K subtilis
%K gene
%K labeling
%K protein
%K fluorescent
%K 作用
%K 生物学
%K 细胞外
%K 蛋白分布
%K 生长缓慢
%K 平行排列
%K 细胞膜
%K 荧光检测
%K 翻译起始位点
%K 序列
%K 结果
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=AE889F374E4D89507BC32F081AB5596C&yid=67289AFF6305E306&vid=B6DA1AC076E37400&iid=B31275AF3241DB2D&sid=C81F81170838C444&eid=07034C6B9EA4A53C&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=16