Salmonella enterica serovar Typhimurium 14028 Suşunda MisL Ototransporter Proteininin İn Vitro Üretimi [1]

Salmonella enterica serovar Typhimurium genomunda kodlanan MisL ototransporter proteini, enfekte konakçı sisteminde eksprese edilmekte, ancak bu bakterinin laboratuvar kültürlerinde üretilmemektedir. Bu çalışmada, MisL ototransporter proteininin in vitro koşullarda üretimi; misL geninin transkripsiyonunu kontrol eden ve pozitif bir transkripsiyonel regülatör olan marT geninin, pBAD24 vektörüne klonlanması ve marT geni bloke edilmiş (?marT) S. Typhimurium 14028 suşuna aktarımı yolu ile gerçekleştirildi. LacZYA: :misL füzyonu içeren rekombinant 14028 suşunda MisL üretimi, ß-galaktozidaz aktivite testi (460 Miller ünitesi) ve Western blot yöntemi kullanılarak tanımlandı. İmmun elektron mikroskobi çalışmaları sonucunda, MisL proteininin bu bakterinin dış yüzeyinde lokalize olduğu saptandı. Bu bulgular MisL ototransporter proteininin S. Typhimurium'un hücre dışı bir matriks adhezini olduğuna işaret etmektedir.

In Vitro Production of MisL Autotransporter Protein in Salmonella enterica serovar Typhimurium 14028

The MisL autotransporter protein is encoded in the genome of the Salmonella enterica serovar Typhimurium, is expressed within an infected host system but not the laboratory cultures of this bacterium. In this study, in vitro production of MisL autotransporter protein was produced by cloning of marT gene, a positive transcriptional regulator controlling expression of misL gene, to the pBAD24 vector and by transforming to marT gene blocked (?marT) S. Typhimurium strain 14028. MisL production in S. Typhimurium 14028 containing lacZYA: misL fusion was determined using both ß-galactosidase activity test (460 Miller units) and Western blotting. Results of immunelectron microscopy studies showed that MisL was localized to outer surface of this bacterium. These data pointed out that MisL is an extracellular matrix adhesin of S. Typhimurium.

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