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    • 21. 发明公开
    • 이스트 GAL80 결핍주를 이용한 재조합 단백질의 생산방법
    • 使用GAL80缺乏的YEAST菌株生产重组蛋白的方法
    • KR1020090016364A
    • 2009-02-13
    • KR1020070080946
    • 2007-08-10
    • 한국생명공학연구원
    • 최의성손여진
    • C12N15/81C12N15/09C12N1/19
    • A GAL80 deficient yeast strain is provided to improve the expression rate and cell growth as compared to the conventional GAL1 deficient strain and eliminate need of galactose which is essential expensive substrate in the cultivation of GAL deficient strain, thereby producing the recombinant protein with low cost and high concentration cell culture. The method for producing a recombinant protein comprises the steps of: crushing the GAL80 gene of yeast; transforming the GAL80 gene crushed yeast strain with the recombinant expression vector encoding the foreign protein; and culturing the transformed yeast stain, wherein the yeast is Saccharomyces cerevisiae.
    • 提供了GAL80缺陷型酵母菌株,以提高与常规GAL1缺陷菌株相比的表达率和细胞生长,并消除了在培养GAL缺陷菌株中必不可少的底物的半乳糖的需要,从而以低成本制备重组蛋白质, 高浓度细胞培养。 制备重组蛋白的方法包括以下步骤:粉碎酵母的GAL80基因; 用编码外源蛋白的重组表达载体转化GAL80基因破碎的酵母菌株; 并培养转化的酵母染色剂,其中酵母是酿酒酵母(Saccharomyces cerevisiae)。
    • 28. 发明公开
    • HARS36서열을 이용하여 효모 한세눌라 폴리모르파균주에서 라이브러리를 제조하는 방법 및 상기 방법에의하여 제조된 라이브러리
    • 使用HARS36序列和由其构建的图书馆在汉森多聚体中构建图书馆的方法
    • KR1020050029374A
    • 2005-03-28
    • KR1020030065622
    • 2003-09-22
    • 한국생명공학연구원
    • 최의성손정훈김소영배정훈
    • C12N15/81
    • C12N15/1024C12N15/815
    • A method for construction of a library in Hansenula polymorpha using a HARS36 sequence and the library constructed by the same method are provided, thereby constructing a useful gene library having high transformation efficiency and uniformed copy number in Hansenula polymorpha without Escherichia coli. The method for construction of the library in Hansenula polymorpha using the HARS36 sequence comprises the steps of: (1) preparing an insertion gene fragment comprising the HARS36 sequence having the nucleotide sequence of SEQ ID NO:1 of which total or portion sequence is exposed to the 5'- or 3'-terminal of the insertion gene fragment and a target expression gene; (2) a receiving gene fragment comprising a selection marker gene, a promoter for the selection marker gene, and the HARS36 sequence having the nucleotide sequence of SEQ ID NO:1 of which total or portion sequence is exposed to the 5'- or 3'-terminal of the receiving gene fragment; (3) transforming Hansenula polymorpha with both gene fragments of steps (1) and (2) to prepare a transformant; and (4) culturing the transformant.
    • 提供了使用HARS36序列构建汉逊酵母(Hansenula polymorpha)的文库的方法以及通过相同方法构建的文库,由此构建了不含大肠埃希氏菌的多形汉逊酵母中具有高转化效率和均匀拷贝数的有用的基因文库。 使用HARS36序列构建多形汉逊酵母的文库的方法包括以下步骤:(1)制备插入基因片段,其包含具有序列号1的核苷酸序列的HARS36序列,其全部或部分序列暴露于 插入基因片段的5'-或3'端和靶表达基因; (2)包含选择标记基因,选择标记基因的启动子和具有SEQ ID NO:1的核苷酸序列的HARS36序列的接收基因片段,其全部或部分序列暴露于5'-或3 接收基因片段的末端; (3)用步骤(1)和(2)的两个基因片段转化多形汉逊酵母以制备转化体; 和(4)培养转化体。
    • 29. 发明授权
    • 한세눌라 폴리모르파 유전자 및 이들 유전자가 결핍된 균주
    • Hansenulla polymorpha基因和这些基因缺陷的菌株
    • KR100354344B1
    • 2002-09-27
    • KR1020020032933
    • 2002-06-12
    • 한국생명공학연구원
    • 최의성이상기손정훈강현아배정훈
    • C12N15/57
    • 본발명은한세눌라폴리모르파유전자및 이들유전자가결핍된균주의제조에있어서, 한세눌라폴리모르파()DL1 유래의카르복시펩티다아제 Y를코딩하는유전자프로티아제 A를코딩하는유전자유전자및 카르복시펩티다아제α를코딩하는유전자을클로닝한다음이 유전자들중,과유전자를유전자에의해디스럽션(disruption)시켜제작한벡터로형질전환된한세눌라폴리모르파 UR2 카르복시펩티다아제 Y, 프로티아제 A 변이주를제조하고또유전자를한세눌라폴리모르파 URA3 유전자팝아웃카세트(pop-out cassette)에의해디스럽션시켜제작한벡터로형질전환된한세눌라폴리모르파 DL1 카르복시펩티다아제 Y, 프로티아제 A, 카르복시펩티다아제α변이주와중복변이주에관한것으로, 상기제조한균주들은외래단백질발현시숙주세포로사용하여카르복시말단분해가감소된외래단백질을제조하는효과가있고또한한세눌라폴리모르파유래의분비시그널펩티드로서사용할수 있는뛰어난효과가있다.
    • 本发明涉及编码编码来源于多形汉逊酵母(Hansenula poly-morpha)(DL1)基因的羧肽酶Y的基因的基因和编码羧肽酶 yujeonjaeul编码α克隆,然后生产转基因世纪克罗纳拉聚莫里帕斯UR2羧基肽酶Y,亲噻唑剂的突变株与由该基因的基因的,gwayu e。通过图(破坏)上午二产生的矢量的 并且还用于与由汉萨克罗纳拉聚莫里帕斯DL1羧基肽酶Y上插图上午二制备的载体转化的世纪克罗纳拉聚莫里帕斯URA3基因弹出盒(弹出输出盒)的基因,亲噻唑要求分组,羧基肽酶 α突变株和重复突变株,将这样制备的菌株用作外源蛋白表达的宿主细胞, 的效果,并且还具有可以用作世纪克罗纳拉聚莫里帕斯衍生的分泌信号肽的优异效果。
    • 30. 发明公开
    • 고농도의 아스타산틴을 생산하는 돌연변이 균주 파피아로도지마 및 이 균주의 발효 배양 방법
    • 突变菌株,PHAFFIA RHODOZYMA(KCTC-0920B)生产酪氨酸及其分离方法
    • KR1020010044210A
    • 2001-06-05
    • KR1020010001721
    • 2001-01-12
    • 해태제과식품주식회사한국생명공학연구원
    • 한지영이승재정명교정광호최의성손정훈심동섭
    • C12N1/16
    • PURPOSE: Provided is a method for separating mutant strain, Phaffia rhodozyma (KCTC-0920B) which generates astaxanthin in a high yield in a short period of time without being degenerated into a wild type during continuous cultivations. CONSTITUTION: The method for separating mutant strain, Phaffia rhodozyma (KCTC-0920B) comprises the steps of: i) cultivating Phaffia rhodozyma (ATCC-96594) in YM broth and treating the strain with 1-methyl-3-nitro-1-nitrosoguanidine (NTG) to kill 98% of yeast; ii) spreading the remaining yeast on YM medium for cultivation and selecting colonies which are growing faster with red color than the original strain; iii) repeating these processes several times and then, cultivating selected bacteria in YM broth for 4-6 days; and iv) measuring the amount of mycobiont and carotenoid and selecting the strain which produces carotenoid in a high yield.
    • 目的:提供一种在短时间内分离产生虾青素的突变菌株红酵母(KCTC-0920B)的方法,而不会在连续培养期间将其变性为野生型。 构成:分离突变菌株,红发夫酵母(KCTC-0920B)的方法包括以下步骤:i)在YM肉汤中培养红发夫酵母(ATCC-96594),并用1-甲基-3-硝基-1-亚硝基胍 (NTG)杀死98%的酵母; ii)将剩余的酵母铺在YM培养基上进行培养,并选择比原始菌株红色生长得更快的菌落; iii)多次重复这些过程,然后在YM肉汤中培养选定的细菌4-6天; 和iv)测量分枝杆菌和类胡萝卜素的量,并以高产率选择产生类胡萝卜素的菌株。