원문정보
유전자 재조합 대장균을 사용한 Alpha-Interferon의 생산과 분비: 제3부: Interferon생산을 위한 유전자의 발현
초록
영어
Alpha-interferon was produced by using recombinant Escherichia coli strains, which carry cloned alpha-interferon gene in plasmid vectors, pIF-III-B and pIF-III-C. With the aid of signal sequence of E. coli lipoprotein, which is placed right in front of the upstream of the cloned alpha-interferon gene of the plasmids, about 50% of alpha-interferon produced was excreted or secreted. Meanwhile, there was no extracellular production of alpha-interferon from the recombinant strain carrying the plasmid Hif-2h that lacks the signal sequence of lipoprotein.
한국어
대장균의 lipoprotein promoter, lac UV5 promoter 및 operator와 lipoprotein의 signal sequence를 가지는 vector에 alpha-IFN유전자를 cloning함으로써 제작된 plasmid plF-III-B와 plF-III-C를 여러종류의 대장균 숙주 세포에 형질 전환하여 IFN의 생산성을 조사해 본 결과 lpp-형질을 가지는 JE5505가 가장 우수했으며, 기존 plasmid, Hif-2h에 비해 130배 높은 생산성을 보였다. 또한 생산된 IFN의 약 50%가 세포 외부로 분비됨을 확인하였다.
목차
INTRODUCTION
MATERIALS AND METHODS
Bacterial strains, plasmids and culture conditions
Induction of gene expression
Transformation
Determination of alpha-IFN activity
Preparation of crude E. coli extracts for the assay of intracellular alpha-lFN
Preparation of samples for the assay of excreted alpha-lFN
Preparation of samples for the assay of alpha-lFN from Periplasmic space
Preparation of standard alpha-IFN solution
RESULTS AND DISCUSSION
Determination of optimal dilution ratio of VSV concentrate for the assay of alpha-lFN activity
Production of alpha-lFN in various E. coli strains harboring various recombinant plasmid
ABBREVIATIONS
요약
REFERENCES