公开(公告)号：KR100411238B1

公开(公告)日：2003-12-18

申请号：KR1020000048124

申请日：2000-08-19

Applicant: 한국과학기술원

Inventor： 변시명 ,  이상준 ,  김동민 ,  배광희 ,  정재훈 ,  옹쉬람

IPC: C12N1/20

Abstract: PURPOSE: Provided is a novel Bacillus sp. whose wprA gene encoding WprA, proteinase of the Bacillus subtilis, is destroyed by homologous recombination to stably express foreign proteins without damaging them. The foreign proteins are produced by transforming the strains with an expression vector and cultured. CONSTITUTION: The preparation method of the Bacillus sp. strain comprises the steps of: constructing gene replacement vector containing wprA gene, Bacillus origin and E. coli origin in which resistant genes to ampicillin and tetracycline are inserted; transforming Bacillus subtilis WB600 using the vector; selecting tetracycline resistant strains and heat-treating them to eliminate the unincorporated vector; screening the genomic DNA of the selected strains by polymerase chain reaction and confirming that wrpA gene is destructed from the genomic DNA of the selected strains; and identifying the strain Bacillus subtilis LB700 (KCTC 18039P) and using it as a host strain to express staphylokinase in a large amount.

Abstract translation: 目的：提供了一种新型芽孢杆菌属。 其编码枯草芽孢杆菌蛋白酶WprA的wprA基因被同源重组破坏，从而稳定表达外源蛋白而不损害它们。 通过用表达载体转化菌株并培养来产生外源蛋白质。 组成：芽孢杆菌的制备方法。 菌株包括以下步骤：构建含有wprA基因，芽孢杆菌来源和大肠杆菌来源的基因置换载体，其中插入对氨苄青霉素和四环素的抗性基因; 使用该载体转化枯草芽孢杆菌WB600; 选择四环素耐药菌株并对它们进行热处理以消除未结合的载体; 通过聚合酶链式反应筛选所选菌株的基因组DNA，并证实wrpA基因从所选菌株的基因组DNA中被破坏; 并鉴定菌株枯草芽孢杆菌LB700（KCTC 18039P）并将其用作宿主菌株以大量表达葡萄球菌激酶。

公开(公告)号：KR1020020014923A

公开(公告)日：2002-02-27

申请号：KR1020000048124

申请日：2000-08-19

Applicant: 한국과학기술원

Inventor： 변시명 ,  이상준 ,  김동민 ,  배광희 ,  정재훈 ,  옹쉬람

IPC: C12N1/20

Abstract: PURPOSE: Provided is a novel Bacillus sp. whose wprA gene encoding WprA, proteinase of the Bacillus subtilis, is destroyed by homologous recombination to stably express foreign proteins without damaging them. The foreign proteins are produced by transforming the strains with an expression vector and cultured. CONSTITUTION: The preparation method of the Bacillus sp. strain comprises the steps of: constructing gene replacement vector containing wprA gene, Bacillus origin and E. coli origin in which resistant genes to ampicillin and tetracycline are inserted; transforming Bacillus subtilis WB600 using the vector; selecting tetracycline resistant strains and heat-treating them to eliminate the unincorporated vector; screening the genomic DNA of the selected strains by polymerase chain reaction and confirming that wrpA gene is destructed from the genomic DNA of the selected strains; and identifying the strain Bacillus subtilis LB700 (KCTC 18039P) and using it as a host strain to express staphylokinase in a large amount.

Abstract translation: 目的：提供一种新型芽孢杆菌 其编码WprA的wprA基因，枯草芽孢杆菌的蛋白酶被同源重组破坏以稳定表达外源蛋白质而不损害它们。 通过用表达载体转化菌株并培养来产生外源蛋白质。 构成：芽孢杆菌的制备方法 菌株包括以下步骤：构建含有wprA基因，芽孢杆菌来源和大肠杆菌来源的基因置换载体，其中插入氨苄青霉素和四环素的抗性基因; 使用载体转化枯草芽孢杆菌WB600; 选择四环素抗性菌株并进行热处理以消除未掺入的载体; 通过聚合酶链反应筛选所选菌株的基因组DNA，并证实wrpA基因从所选菌株的基因组DNA中破坏; 并鉴定菌株枯草芽孢杆菌LB700（KCTC 18039P），并将其用作宿主菌株以大量表达葡萄球菌激酶。