원문정보
전기장 충격법에 의한 코리네헝 세균의 고효율 헝질전환
초록
영어
Escherkchla coli/Cownebacterium glutamicum shuttle vectors, pECCGl and pECCG2 were constructed by joining a 3.0 kb C. glutamicum cryptic plasmid pCBl and a 3.94 kb E. coli plasmid pACYC177. Using the plasmid pECCGl, various parameters involved in electroporation system including electric field strength, resistance, DNA concentration, cell concentration and growth stage were investigated independently and optimized for the high efficiency transformation of coryneform bacteria. Transformation efficiencies of 106 transformants/ug of plasmid DNA were achieved with Corynebacterium glutamicum.
한국어
대장균과 코리네형 세균간의 shutle vector pECCGI과 pECCD2를 제작하고, plasmid pECCGI과 glycine배지에서 다양한 Corynebacterium glutamicum을 사용하여 전기장 충격법에 의한 형질전환에 있어서 여러 조건을 조사한 결과 세포 현탁액 40ul와 DNA 2ul의 혼합액 사용시 저항 600 obms, 전기장의 세기 12.5kv/cm, DNA양 10ng, 세포수 $4.510^8$와 세포회수 시기를 1.0이하의 A600으로 했을때 106 transformants/ug of DNA의 형질전환 효율을 보였다
목차
INTRODUCTION
MATERIALS AND METHODS
Bacterial strains and plasmids
Media and bacterial cultivation
Plasmid DNA preparations and DNA manipulations
Electroporation equipment and electroporation procedure
RESULTS
Construction of E. coli/C. glutamicum shuttle vectors
Protoplase transformation of C. glutamicum
Optimization of conditions for C. glutamicum electroporation
Resistance
Electric field strength
DNA concentration
Cell concentration
Cell harvest stage
Effect of penicilin-G in the culture medium
DISCUSSION
ABBREVIATIONS
요약
REFERENCES