公开(公告)号：KR100253546B1

公开(公告)日：2000-04-15

申请号：KR1019970059686

申请日：1997-11-13

Applicant: 강봉균

Inventor： 김형규 ,  강봉균

IPC: C12N15/12 ,  C12N15/62

Abstract: PURPOSE: Provided is a green fluorescence protein (GFP) mutant which is very useful as a reporter molecule to monitor gene expression and protein position while holding bioluminescence and clarifying domains. CONSTITUTION: A method for manufacturing a green fluorescence protein (GFP) mutant is characterized by the following steps of: i) amplifying NEX δ-GFP segments that code GFP through polymerase chain reaction (PCR) using primers, gfp1 (5'GGGAATTCCATATGAGAAAAGGAGAAGA) and gfp2 (5'GACACCAGACCAACTGG); ii) cutting PCR products with restriction enzymes Nde I and BamH I and inserting vector Nde I/BamH I-linear pET3a inducing GFP into E. coli; and iii) manufacturing various GFP mutants by using restriction enzymes, exonuclease, and recombinant primers. And the strength of bioluminescence of GFP mutants are measured in E. coli transformed by pET3a-GFP.

Abstract translation: 目的：提供绿色荧光蛋白（GFP）突变体，作为报告分子非常有用，用于监测基因表达和蛋白质位置，同时保持生物发光和澄清结构域。 构成：用于制备绿色荧光蛋白（GFP）突变体的方法的特征在于以下步骤：i）使用引物gfp1（5'GGGAATTCCATATGAGAAAAGGAGAAGA）和通过聚合酶链反应（PCR）扩增编码GFP的NEXδ-GFP片段， gfp2（5'GACACCAGACCAACTGG）; ii）用限制酶Nde I和BamH I切割PCR产物，并将导入GFP的载体Nde I / BamH I线性pET3a插入大肠杆菌中; 和iii）通过使用限制酶，核酸外切酶和重组引物制备各种GFP突变体。 并且在通过pET3a-GFP转化的大肠杆菌中测量GFP突变体的生物发光强度。

公开(公告)号：KR1019990039547A

公开(公告)日：1999-06-05

申请号：KR1019970059686

申请日：1997-11-13

Applicant: 강봉균

Inventor： 김형규 ,  강봉균

IPC: C12N15/12 ,  C12N15/62

Abstract: 젤리피쉬 아에쿠오레아 빅토리아의 녹색 형광 단백질(GFP) 및 그 돌연변이 S65T 및 I167T의 C-말단으로부터 7~9개의 아미노산 성분의 절단은 형광 활성을 파괴하지 않고, 특히 9개의 아미노산 성분이 절단된 GFP 결실 돌연변이들은 더욱 강력한 형광 활성을 나타냄으로써 유전자 발현 및 단백질 위치의 모니터링(monitoring)을 위한 리포터(reporter) 분자로 유용하게 사용될 수 있다.