This research was carried out for the purpose of developing shuttle vectors which can express foreigns genes efficiently in both $\underline{Zymomonas}$ $\underline{mobilis}$and $\underline{Eschericha}$ $\underline{coli}$. The initial approach was done by isolating and charaterizing $\underline{Z.}$ $\underline{mobilis}$ DNA fragments having promoter activity in $\underline{E.}$ $\underline{coli}$. For this, a promoter screening vector, pCMT215 was constructed by ligating pMT21 with chloramphenicol acetyltransferase (CAT) gene from pYEJ001. The pMT21 is a plasmid containing -lactamase gene and multiple cloning sites without nonspecific binding with $\underline{E.}$ $\underline{coli}$ RNA polymerase. A promoterless CAT structural gene was available from pYEJ001. Fragments of $\underline{Z.}$ $\underline{mobilis}$ DNA were inserted in pCMT215 and a gene library was prepared in $\underline{E.}$ $\underline{coli}$. Fourteen clones out of this library showed resistance to chloramphenicol ranging from 30 ug/ml to 750 ug/ml of broth. The recombinant plasmids isolated from the Cm transformants contained inserts with the size of 170-1800 base pairs. Among fourteen clones, five clones were selected and their nucleotide sequencies were analysed and their bases for transcription initiation were determined. The $\underline{Z.}$ $\underline{mobilis}$ DNA fragments examined share many features described for the promoter regions such as partial sequence homology with -35 and -10 region of $\underline{E.}$ $\underline{coli}$ consensus promoters, AT rich regions, poly A's or T's stretches and palindromic sequences. The positions of transcriptional initiation appeared at more than one site spaced over by 4 to 170 base pairs in $\underline{E.}$ $\underline{coli}$. All the fourteen clones have a potential to be developed as a shuttle expression vector. Basic informations on the requirements in sequences of the transcription and translation in $\underline{Z.}$ $\underline{mobilis}$ will be also available.
$\underline{Eschericha}$ $\underline{coli}$ 내에서 프로모터 활성을 보이는 $\underline{Zymomonas}$ $\underline{mobilis}$ 유전자 절편을 분리하고 특성을 분석하였다. 프로모터 탐색용 백터인 pCMT215는 pYEJ001 내부의 클로람페니콜 아세틸 전이 효소 유전자와 pMT21을 접합시켜 조립하였다. 모두 14개의 형질전환주가 클로람페니콜에 내성을 보였으며, 내성정도는 배지 1ml당 30 - 750ug 이었다. 클로닝된 유전자 조각의 크기는 0.1 - 1.5 kb였다. 그중 5개의 유전자 조각의 염기서열을 분석해본 결과 프로모터 지역에서 발견되는 일반적인 특성, $\underline{E.}$ $\underline{coli}$ 프로모터의 -35 또는 -10 지역과의 부분적 일치, AT 염기가 풍부한 지역, 연속적인 A 또는 T 염기, 회문형태의 염기서열등이 발견 되었다. 또한 프라이머 연장 실험 결과, 전사의 시작이 4 - 170 염기의 거리를 두고 두 곳 또는 여러곳에서 일어남을 볼 수 있었다.
