公开(公告)号：US5707828A

公开(公告)日：1998-01-13

申请号：US433037

申请日：1995-05-03

申请人： Kotikanyadan Sreekrishna ,  Juerg F. Tschopp ,  Gregory P. Thill ,  Russell A. Brierley ,  Kathryn A. Barr

发明人： Kotikanyadan Sreekrishna ,  Juerg F. Tschopp ,  Gregory P. Thill ,  Russell A. Brierley ,  Kathryn A. Barr

IPC分类号： C12N15/09 ,  C07K14/765 ,  C12N1/15 ,  C12N1/19 ,  C12N15/14 ,  C12N15/81 ,  C12P21/02 ,  C12R1/84 ,  C12P21/06 ,  C12N15/00 ,  C12N15/63

CPC分类号： C12N15/815 ,  C07K14/765

摘要： The present invention is directed to expression cassettes comprising a 5' regulatory region from at least one of the Pichia pastoris AOXI gene, p40 gene, DASI gene or HIS4 gene, operably linked to an HSA structural gene including the HSA signal sequence. The HSA structural gene has a translational start codon within 0 to 11 deoxyribonucleotides from the 5' end of the HSA structural gene and is operably linked to a 3' termination sequence. Further, the adenine and thymine content of the intervening deoxyribonucleotides is in the range from about 55 to about 64%. The expression cassette may be an autonomously replicating vector or an integrative vector. Other embodiments of the present invention include Pichia pastoris strains transformed with the HSA expression cassettes and processes for secretion of HSA using the expression cassettes.

摘要翻译： 本发明涉及包含与包含HSA信号序列的HSA结构基因可操作地连接的至少一种巴斯德毕赤酵母AOXI基因，p40基因，DASI基因或HIS4基因的5'调节区的表达盒。 HSA结构基因具有来自HSA结构基因5'末端的0至11个脱氧核糖核苷酸之间的翻译起始密码子，并且可操作地连接到3'终止序列。 此外，介入的脱氧核糖核苷酸的腺嘌呤和胸腺嘧啶含量在约55至约64％的范围内。 表达盒可以是自主复制载体或整合载体。 本发明的其他实施方案包括用HSA表达盒转化的巴斯德毕赤酵母菌株和使用表达盒分泌HSA的方法。

公开(公告)号：US5002876A

公开(公告)日：1991-03-26

申请号：US909528

申请日：1986-09-22

申请人： Kotikanyadan Sreekrishna ,  Motohiro Fuke ,  Rica H. Potenz

发明人： Kotikanyadan Sreekrishna ,  Motohiro Fuke ,  Rica H. Potenz

IPC分类号： C12N15/09 ,  C07K14/525 ,  C12N1/16 ,  C12N15/12 ,  C12N15/68 ,  C12N15/81 ,  C12P21/00 ,  C12P21/02 ,  C12R1/84

CPC分类号： C12N15/815 ,  C07K14/525 ,  C12N15/68

摘要： Novel DNA constructs comprising yeast regulatory regions plus the structural coding region for human tumor necrosis factor, are disclosed. These novel constructs are incorporated into a variety of linear and circular plasmids. Such plasmids are used for yeast transformation and ultimately for the production of human tumor necrosis factor by yeast.

摘要翻译： 公开了包含酵母调节区加上人肿瘤坏死因子结构编码区的新型DNA构建体。 这些新型构建体并入多种线性和环状质粒。 这些质粒用于酵母转化，最终用于通过酵母生产人肿瘤坏死因子。

公开(公告)号：US5330901A

公开(公告)日：1994-07-19

申请号：US691079

申请日：1991-04-25

申请人： William D. Prevatt ,  Kotikanyadan Sreekrishna

发明人： William D. Prevatt ,  Kotikanyadan Sreekrishna

IPC分类号： C12N15/09 ,  C07K14/765 ,  C12N1/19 ,  C12N1/32 ,  C12N15/14 ,  C12N15/81 ,  C12P21/02 ,  C12R1/84

CPC分类号： C12N15/815 ,  C07K14/765 ,  C12N1/32 ,  C07K2319/00 ,  C07K2319/036

摘要： A process for the production of HSA in Pichia pastoris cells comprising cultivating Pichia pastoris cells capable of expressing HSA at a pH of about 5.7 to about 6.4 contemporaneously with the expression of HSA.

摘要翻译： 一种用于在巴斯德毕赤酵母细胞中产生HSA的方法，其包括与HSA的表达同时培养能够在约5.7至约6.4的pH下表达HSA的巴斯德毕赤酵母细胞。

公开(公告)号：US4857467A

公开(公告)日：1989-08-15

申请号：US887300

申请日：1986-07-23

申请人： Kotikanyadan Sreekrishna ,  Motohiro Fuke

发明人： Kotikanyadan Sreekrishna ,  Motohiro Fuke

IPC分类号： C12N9/26 ,  C12N15/65 ,  C12N15/81

CPC分类号： C12N9/2408 ,  C12N15/65 ,  C12N15/81 ,  C12N15/815 ,  Y10S435/938

摘要： Yeast strains are transformed with, and selected for, using DNA fragments encoding a gene function not present in the wild-type yeast strains. The invention requires transforming the yeast with DNA fragments which comprise the gene function which the wild-type yeast strain is lacking; regeneration of the transformants on a non-selective regeneration medium; then selection of those colonies which are capable of growth on a carbon and energy source which requires expression of the gene function provided by the DNA fragment. Thus, transformation with such DNA fragments allows for positive selection of transformed strains.

摘要翻译： 酵母菌株用编码野生型酵母菌株中不存在的基因功能的DNA片段转化并选择。 本发明需要用包含野生型酵母菌株缺乏的基因功能的DNA片段来转化酵母; 在非选择性再生培养基上再生转化体; 然后选择能够在需要表达由DNA片段提供的基因功能的碳和能量源上生长的那些菌落。 因此，用这种DNA片段的转化允许转化菌株的阳性选择。