Expression and purification of recombinant proteins by fusion to maltose-binding protein

被引:89
作者
Riggs, P [1 ]
机构
[1] New England Biolabs Inc, Beverly, MA 01915 USA
关键词
primer; restriction; genomic DNA; viral RNA; PCR; RT-PCR; amplification;
D O I
10.1385/MB:15:1:51
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
pMAL vectors provide a method for purifying proteins from cloned genes bp fusing them to maltose-binding protein (MBP, product of malE), which hinds to amylose. The vectors use the tac promoter and the translation initiation signals of MBP to give high-level expression of the fusion, and an affinity purification for MBP to isolate the fusion protein. The pMAL polylinkers carry restriction sites to insert the gene of interest and encode a site for a specific protease to separate MBP from the target protein after purification. Vectors with or without the malE signal sequence can be used, to express the protein cytoplasmically for the highest level of production or periplasmically tct help in proper folding of disulfide-bonded proteins.
引用
收藏
页码:51 / 63
页数:13
相关论文
共 21 条
[1]   ATG VECTORS FOR REGULATED HIGH-LEVEL EXPRESSION OF CLONED GENES IN ESCHERICHIA-COLI [J].
AMANN, E ;
BROSIUS, J .
GENE, 1985, 40 (2-3) :183-190
[2]  
BLOCH K, 1989, CURRENT PROTOCOLS MO
[3]   ENGINEERING SUBTILISIN BPN' FOR SITE-SPECIFIC PROTEOLYSIS [J].
CARTER, P ;
NILSSON, B ;
BURNIER, JP ;
BURDICK, D ;
WELLS, JA .
PROTEINS-STRUCTURE FUNCTION AND GENETICS, 1989, 6 (03) :240-248
[4]   PRODUCTION OF RECOMBINANT BOVINE ENTEROKINASE CATALYTIC SUBUNIT IN ESCHERICHIA-COLI USING THE NOVEL SECRETORY FUSION PARTNER DSBA [J].
COLLINSRACIE, LA ;
MCCOLGAN, JM ;
GRANT, KL ;
DIBLASIOSMITH, EA ;
MCCOY, JM ;
LAVALLIE, ER .
BIO-TECHNOLOGY, 1995, 13 (09) :982-987
[5]  
DUPLAY P, 1984, J BIOL CHEM, V259, P606
[6]   CLEAVAGE AND PURIFICATION OF PROKARYOTICALLY EXPRESSED HIV GAG AND ENV FUSION PROTEINS FOR DETECTION OF HIV ANTIBODIES IN THE ELISA [J].
ELLINGER, S ;
MACH, M ;
KORN, K ;
JAHN, G .
VIROLOGY, 1991, 180 (02) :811-813
[7]  
GUAN C, 1988, GENE, V67, P21
[8]  
JOHNSTON TC, 1986, J BIOL CHEM, V261, P4805
[9]  
KELLERMANN OK, 1982, METHOD ENZYMOL, V90, P459
[10]  
Lauritzen C, 1991, Protein Expr Purif, V2, P372, DOI 10.1016/1046-5928(91)90096-2