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1. 发明授权

KR101508635B1 유전체 수준의 무작위 돌연변이 유도기술 有权
标题翻译：全基因组随机诱变方法
公开(公告)号：KR101508635B1
公开(公告)日：2015-04-07
申请号：KR1020130097463
申请日：2013-08-16
申请人： 한국생명공학연구원
发明人： 권오석 , 오두병 , 김오철 , 황동현
IPC分类号： C12N1/19 , C12N15/52 , C12N15/80
摘要： 본발명은교정기능이결손된변이 DNA 중합효소델타의촉매소단위(DNA polymerase delta catalytic subunit) Pol3를이용한한세눌라폴리모파게놈의무작위돌연변이유도기술에관한것으로, 구체적으로교정기능이결손된 Pol3 단백질을생산하는신규한발현벡터를한세눌라폴리모파에도입후, 표적형질을나타내는돌연변이균주를분리동정하고, 더이상의돌연변이축적을방지하기위하여본 발명의교정기능이결손된 Pol3 단백질발현벡터를제거하는방법을확립하였다. 본발명의무작위돌연변이유도기술은산업상유용한균주로알려진한세눌라폴리모파의알려지지않거나복합적인돌연변이를유도하여표적형질을나타내는돌연변이를제조하는데있어서유용하게사용될수 있다.

2. 发明授权

KR101419214B1 LacI 단백질 변이체를 이용한 독성 단백질의 생산 방법 有权
标题翻译：使用LACI蛋白质突变体生产毒性蛋白的方法
公开(公告)号：KR101419214B1
公开(公告)日：2014-07-14
申请号：KR1020130002227
申请日：2013-01-08
申请人： 한국생명공학연구원
发明人： 윤성호 , 이대희 , 김성근 , 김오철
IPC分类号： C12N15/63 , C12N15/70
摘要： The present invention relates to a recombinant vector including a Rha promoter derived from colon bacillus, and a gene for coding a LacI (lac repressor) protein mutant whose at least one amino acid is mutated among 75th, 192th, 197th, or 225th amino acids of LacI (lac repressor) protein consisting of an amino acid sequence of a sequence number one connected with the promoter to be able to operate; and a method for producing toxic protein comprising a step for expressing a gene by transforming a host cell after inserting the gene for coding toxic protein and the host cell transformed into the recombinant vector into a recombinant expression vector. The expression of a LacI mutant gene is precisely controlled rhamnose concentration-dependently by using a Rha promoter controlled by rhamnose, so the expression of a target protein to be produced inside a colon bacillus cell is optimized and the output is increased. Therefore, it is useful for the expression of the recombinant toxic protein in a host cell including colon bacillus.
摘要翻译：本发明涉及包含来自大肠杆菌的Rha启动子的重组载体和用于编码LacI（lac阻遏物）蛋白质突变体的基因，其在至少一个氨基酸在第75,192,197或225个氨基酸中突变 LacI（lac阻遏物）蛋白，其由与能够操作的启动子连接的序列号的氨基酸序列组成; 以及生产有毒蛋白质的方法，包括将插入用于编码毒性蛋白质的基因和转化到重组载体中的宿主细胞插入重组表达载体后，通过转化宿主细胞来表达基因的步骤。 LacI突变基因的表达通过使用由鼠李糖控制的Rha启动子来精确控制鼠李糖浓度，因此优化在大肠杆菌细胞内产生的靶蛋白的表达，并且输出增加。因此，对包含大肠杆菌的宿主细胞中的重组毒性蛋白质的表达是有用的。

3. 发明公开

KR1020140047520A 유전체 수준의 무작위 돌연변이 유도기술 有权
标题翻译：广义随机变形的方法
公开(公告)号：KR1020140047520A
公开(公告)日：2014-04-22
申请号：KR1020130097463
申请日：2013-08-16
申请人： 한국생명공학연구원
发明人： 권오석 , 오두병 , 김오철 , 황동현
IPC分类号： C12N1/19 , C12N15/52 , C12N15/80
摘要： The present invention relates to a technique for inducing a random mutation of Hansenula polymorpha genomes using Pol3 that is a DNA polymerase delta catalytic subunit in which the correcting function is deleted. More particularly, a novel expression vector that generates Pol3 proteins in which the correcting function is deleted is transduced to Hansenula polymorpha and mutant strains representing target traits are isolated and identified, and the Pol3 protein expression vector in which the correcting function is deleted is eliminated so as to prevent the accumulation of mutants. The technique of the present invention can be effectively used to produce mutants representing target traits by inducing unknown or complex mutations of Hansenula polymorpha, which is known as an industrially useful strain.
摘要翻译：本发明涉及一种使用Pol3修饰汉逊酵母多形汉逊酵母基因组的随机突变技术，该Pol3是缺失校正功能的DNA聚合酶δ催化亚单位。更具体地说，分离和鉴定产生其中缺失校正功能的Pol3蛋白质的新型表达载体，并转录到多形汉逊酵母（Hansenula polymorpha），并且鉴别出其中缺失校正功能的Pol3蛋白质表达载体，因此消除以防止突变体的积累。本发明的技术可以有效地用于产生代表目标性状的突变体，通过诱导多形汉逊酵母（Hansenula polymorpha）的未知或复杂的突变，这被称为工业上有用的菌株。

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