Restriction digests of the clone give the following sizes (kb): BamHI/NcoI--5.8, 2.0; EcoRI--6.0, 1.8; HindIII--7.0, 0.6; PstI--6.2, 1.6; PvuII--3.0, 2.8, 1.5, 0.3. Constructed by ligating a PCR product creating an NcoI site on the 5' end to a fragment from a cDNA library. The insert contains the following restriction sites (approximate kb from the 5' end): EcoRI--0.60; HindIII--1.75; NdeI--0.67; PstI--1.55; PvuII--1.31, 1.62; SacI--0.09. Transcription from the T7 promoter allows expression of a kfc-fusion protein that can be purified by affinity chromatography on calmodulin-agarose. (kfc refers to kemptide - factor X - calmodulin-binding domains). |
