不可分型流感嗜血杆菌特异性异位递送载体的构建

Construction of a nontypeable Haemophilus influenzae-specific ectopic delivery vector.

作者信息

Daines Dayle A, Smith Arnold L

机构信息

Seattle Biomedical Research Institute, 4 Nickerson St. Suite 200, Seattle, WA 98109, USA.

出版信息

J Microbiol Methods. 2004 Jun;57(3):421-4. doi: 10.1016/j.mimet.2004.02.014.

DOI:10.1016/j.mimet.2004.02.014

PMID:15134890

Abstract

Complementation of chromosomal mutations in trans can introduce artifacts due to the number of episomal copies of the gene in question. One solution is to study the gene expressed at a single ectopic site in cis. We have designed and constructed a vector that allows homologous recombination into a gene encoding a frame-shifted IS1016-V6 protein in the Haemophilus influenzae Rd KW20 chromosome (HI1018). This site is the location of the > or = 35 kilobase capsule locus in encapsulated type b and d strains. This locus is not present in the nontypeable Rd KW20 strain, thus allowing ectopic expression of genes homologously recombined into HI1018 without polar effects.

摘要

由于所讨论基因的附加体质粒拷贝数，反式互补染色体突变可能会引入假象。一种解决方案是研究在顺式中单个异位位点表达的基因。我们设计并构建了一种载体，该载体允许同源重组到流感嗜血杆菌Rd KW20染色体（HI1018）中编码移码IS1016-V6蛋白的基因中。该位点是b型和d型包膜菌株中≥35千碱基荚膜基因座的位置。该基因座在不可分型的Rd KW20菌株中不存在，因此允许同源重组到HI1018中的基因进行异位表达而无极性效应。