Abstract
A secretion signal sequence-selection vector (pGS40) was constructed based on an α-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore α-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgarno (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.
| Original language | English |
|---|---|
| Pages (from-to) | 1052-1056 |
| Number of pages | 5 |
| Journal | Journal of Microbiology and Biotechnology |
| Volume | 14 |
| Issue number | 5 |
| Publication status | Published - 2004 Oct 1 |
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Keywords
- α-amylase
- Lactococcus lactis ssp. cremoris LM0230
- Secretion signal
- Signal-sequence selection vector
ASJC Scopus subject areas
- Biotechnology
- Applied Microbiology and Biotechnology
- Microbiology
- Bioengineering
Cite this
Screening and characterization of secretion signals from Lactococcus lactis ssp. cremoris LM0230. / Jeong, Do Won; Choi, Youn Chul; Lee, Jung Min; Seo, Jung Min; Kim, Jeong Hwan; Lee, Jong Hoon; Kim, Kyoung Heon; Lee, Hyong Joo.
In: Journal of Microbiology and Biotechnology, Vol. 14, No. 5, 01.10.2004, p. 1052-1056.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Screening and characterization of secretion signals from Lactococcus lactis ssp. cremoris LM0230
AU - Jeong, Do Won
AU - Choi, Youn Chul
AU - Lee, Jung Min
AU - Seo, Jung Min
AU - Kim, Jeong Hwan
AU - Lee, Jong Hoon
AU - Kim, Kyoung Heon
AU - Lee, Hyong Joo
PY - 2004/10/1
Y1 - 2004/10/1
N2 - A secretion signal sequence-selection vector (pGS40) was constructed based on an α-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore α-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgarno (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.
AB - A secretion signal sequence-selection vector (pGS40) was constructed based on an α-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore α-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgarno (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.
KW - α-amylase
KW - Lactococcus lactis ssp. cremoris LM0230
KW - Secretion signal
KW - Signal-sequence selection vector
UR - http://www.scopus.com/inward/record.url?scp=8644289453&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=8644289453&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:8644289453
VL - 14
SP - 1052
EP - 1056
JO - Journal of Microbiology and Biotechnology
JF - Journal of Microbiology and Biotechnology
SN - 1017-7825
IS - 5
ER -