发明名称 |
Bacteriorhodopsin fusion membrane protein expression system |
摘要 |
An expression vector is disclosed, which comprises: a) a polynucleotide sequence encoding a bacteriorhodopsin or a mutant bacteriorhodopsin; b) a multiple cloning site; c) a T7 promoter, d) a polyhistidine tag; e) a first protease cleavage site; f) optionally a second protease cleavage site; and g) optionally a linker; wherein the mutant bacteriorhodopsin comprises the residue corresponding to Asn94 of SEQ ID NO: 1. Also disclosed is a fusion membrane protein expression system, which comprises: a) a polynucleotide sequence encoding a mutant Haloarcula marismortui bacteriorhodopsin/D94N (HmBRI/D94N) or a Haloquadratum walsbyi bacteriorhodopsin (HwBR); b) a target membrane protein; and c) a T7 promoter, operably linked to the mutant HmBRI/D94N or HwBR and the target membrane protein. Host cells comprising the expression vector or the fusion membrane protein expression system and methods of using the same are also disclosed. |
申请公布号 |
US9067985(B2) |
申请公布日期 |
2015.06.30 |
申请号 |
US201314041493 |
申请日期 |
2013.09.30 |
申请人 |
ACADEMIA SINICA |
发明人 |
Wang Andrew H.-J.;Hsu Min-Feng;Yang Chii-Shen;Fu Hsu-Yuang |
分类号 |
C07K14/215 |
主分类号 |
C07K14/215 |
代理机构 |
Intellectual Property Connections, Inc. |
代理人 |
Saunders Hsiu-Ming;Intellectual Property Connections, Inc. |
主权项 |
1. An expression vector comprising:
a) a polynucleotide sequence encoding a mutant bacteriorhodopsin having at least 80% sequence identity to SEQ ID NO: 1; b) a multiple cloning site, located downstream from the polynucleotide encoding the mutant bacteriorhodopsin; c) a T7 promoter, operably linked to the polynucleotide sequence encoding the mutant bacteriorhodopsin, d) a nucleic acid sequence encoding a polyhistidine tag, located downstream from the polynucleotide sequence encoding the mutant bacteriorhodopsin and upstream from the multiple cloning site; e) a first polynucleic acid encoding a first protease cleavage site, located downstream from the nucleic acid sequence encoding the polyhistidine tag and upstream from the multiple cloning site; f) optionally a second polynucleic acid encoding a second protease cleavage site, located between the polynucleotide sequence encoding the mutant bacteriorhodopsin and the nucleic acid sequence encoding the polyhistidine tag; and g) optionally a nucleotide sequence encoding a linker located between the first polynucleic acid encoding the first protease cleavage site and the multiple cloning site;wherein the mutant bacteriorhodopsin retains Asn94 of SEQ ID NO: 1. |
地址 |
Taipei TW |