A pair of ligation-independent Escherichia coli expression vectors for rapid addition of a polyhistidine affinity tag to the N- or C-termini of recombinant proteins.

ABSTRACT: 6x His tag is one of the most widely used affinity fusion tags that facilitates detection and purification of recombinant proteins. However, the location of this tag within a particular type of protein may influence the expression, solubility, and bioactivity of the protein, and the optimal location needs to be determined experimentally. To provide a tool for rapid generation of 6x His tags at the N- or C-terminus of any recombinant protein, we have constructed a pair of Escherichia coli expression vectors-pLIC-NHis and pLIC-CHis-based on the pET30a vector, for ligation-independent cloning (LIC). Construction of this new pair of LIC vectors was accomplished by replacement of the multiple cloning site of pET30a with two specifically designed LIC cloning sites. A target gene derived by PCR w

SUBMITTER: Dan H 

PROVIDER: S-EPMC2777343 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature