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KMID : 0545120070170081316
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 8 p.1316 ~ p.1323
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Development of a Protein Secretion System with the Application of Secdependent Protein Secretion Components
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Kim Sam-Woong
Kim Young-Hee
Yoo Ah-Young
Yu Jong-Earn
Cha Jae-Ho
Hur Jin
Lee John-Hwa
Kang Ho-Young
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Abstract
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In order to induce high levels of protein secretion, we have constructed a recombinant plasmid, designated pBP244, into which was incorporated key components of the type-II Sec-dependent secretion system, including LepB (signal peptidase), SecA (ATPase), and SecB (chaperone). The biological activities of the LepB, SecA, and SecB components expressed from genes harbored by pBP244 appeared to play their normal roles. In order to evaluate the protein secretion, a pspA (Streptococcus pneumoniae surface protein A) gene was cloned into pBP244, resulting in pBP438. S. typhimurium harboring pBP438 was grown until the stationary phase, and secreted a higher level of PspA into the culture supernatants than did the strain harboring pYA3494. The strain harboring pBP438 secreted a supernatant amount 1.71-fold, a periplasmic space amount 1.47-fold, and an outer membrane amount 1.49-fold higher than that of pYA3494. S. typhimurium c8554 kept the Asd+ plasmid pBP244 and pBP438 for 60 generations in LB broth harboring DAP, thereby indicating that pBP244 and pBP438 were quite stable in the Salmonella strain
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KEYWORD
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pBP244, protein secretion, type II Sec-dependent secretion system, LepB, SecA, SecB
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