Boyle-Vavra S, Seifert H S
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, IL 60611, USA.
Gene. 1995 Mar 21;155(1):101-6. doi: 10.1016/0378-1119(94)00890-5.
Shuttle mutagenesis is a method for producing stable mini-transposon (mTn) insertions into the genome of Neisseria gonorrhoeae (gonococcus, Gc) and other microbes. Using an mTn3 derivative, we have produced an mTn (mTnCmPhoA) which contains a phoA' gene lacking its N-terminal signal sequence useful for isolating genes which encode exported proteins. mTnCmPhoA was characterized in Gc and Escherichia coli using a cloned target containing the Gc genes, opaE1, pilA and pilB. PhoA+ Gc containing pilB::mTnCmPhoA insertions confirm that PilB is an exported protein in Gc. This system will be useful for isolating and characterizing extracytoplasmic virulence factors from Gc and other bacterial pathogens.
穿梭诱变是一种将稳定的微型转座子（mTn）插入淋病奈瑟菌（淋球菌，Gc）和其他微生物基因组的方法。利用mTn3衍生物，我们构建了一个mTn（mTnCmPhoA），它包含一个缺失N端信号序列的phoA'基因，该信号序列可用于分离编码分泌蛋白的基因。使用含有Gc基因opaE1、pilA和pilB的克隆靶标，在Gc和大肠杆菌中对mTnCmPhoA进行了表征。含有pilB::mTnCmPhoA插入片段的PhoA+ Gc证实PilB是Gc中的一种分泌蛋白。该系统将有助于从Gc和其他细菌病原体中分离和鉴定胞外毒力因子。
