Document Detail


ThyA as a selection marker in construction of food-grade host-vector and integration systems for Streptococcus thermophilus.
MedLine Citation:
PMID:  15006818     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We constructed food-grade host-vector and integration systems for Streptococcus thermophilus by using a thymidylate synthase gene (thyA) as the selection marker. Two thyA genes, thyA(St) and thyA(Lb), were cloned from S. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, respectively. Thymidine-requiring mutants of S. thermophilus were obtained after successive cultures in the presence of trimethoprim, and one of them, TM1-1, was used as the host. Food-grade vectors were constructed by using either thyA(St) or thyA(Lb) as the selection marker. Transformants of TM1-1 created by using these vectors were selected for thymidine autotrophy as efficiently as for erythromycin resistance. By using the host-vector system developed in this way, a foreign amylase gene (amyA) was expressed in TM1-1 and was also integrated into the chromosome by use of a temperature-sensitive integration vector constructed with thyA(Lb) as the selection marker via a double-crossover event. The results obtained show that thyA is an efficient and safe selection marker for S. thermophilus that is suitable for food applications.
Authors:
Yasuko Sasaki; Yoshiyuki Ito; Takashi Sasaki
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  70     ISSN:  0099-2240     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2004 Mar 
Date Detail:
Created Date:  2004-03-09     Completed Date:  2004-05-19     Revised Date:  2013-04-18    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1858-64     Citation Subset:  IM    
Affiliation:
Food Functionality Research Institute, Meiji Dairies Corporation, Odawara, Kanagawa 250-0862, Japan. YASUKO_SASAKI@MEIJI-MILK.COM
Data Bank Information
Bank Name/Acc. No.:
GENBANK/E12778;  E12779
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
DNA, Bacterial / genetics
Food Microbiology*
Food Technology
Genes, Bacterial
Genetic Markers
Genetic Vectors
Humans
Lactobacillus / enzymology,  genetics
Molecular Sequence Data
Mutagenesis
Restriction Mapping
Streptococcus / enzymology*,  genetics*
Thymidylate Synthase / genetics*
Transformation, Genetic
Yogurt / microbiology
Chemical
Reg. No./Substance:
0/DNA, Bacterial; 0/Genetic Markers; EC 2.1.1.45/Thymidylate Synthase
Comments/Corrections

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