Results: 2

1.
Figure 1

Figure 1. From: Expression of Recombinant Proteins with Uniform N-Termini.

Primary structure of the intein–trypsinogen fusion protein. An initiator methionine was placed upstream of the 154 amino-acid long mini-intein, which was then fused in-frame with the 232 amino-acid long human cationic trypsinogen. At the fusion junction, the C-terminal Asn of the splice region is joined to Ala16 of the trypsinogen propeptide. The N-terminal cysteine of the mini-intein has been mutated to Ala to disable splicing at the N-terminus. The sequence of the fusion construct has been deposited with GenBank under accession number DQ371396. Reproduced from ref. 9 with permission from Elsevier Science.

Orsolya Király, et al. Methods Mol Biol. ;705:175-194.
2.
Figure 2

Figure 2. From: Expression of Recombinant Proteins with Uniform N-Termini.

Expression and purification of the intein–trypsinogen fusions. In the experiment shown here, wild type cationic trypsinogen and pancreatitis-associated mutants p.A16V and p.N29I were expressed and purified. SDS–PAGE analysis of (A) inclusion bodies from LG-3 cells expressing the intein–trypsinogen fusions; (B) trypsinogens eluted from the ecotin-affinity column; (C) trypsinogen fractions after Mono S chromatography. Inclusion body fraction prepared from 1.8 mL LG-3 culture (OD600 = 1.6) or ~5 μg purified protein was loaded per lane. (D) Mono S chromatography of cationic trypsinogen eluted from the ecotin-affinity column. The column was developed with a 0–0.5M gradient of NaCl at a flow rate of 1mL/min. Under these conditions only pure trypsinogen is eluted, while unprocessed fusion proteins are not recovered. Reproduced from ref. 9 with permission from Elsevier Science.

Orsolya Király, et al. Methods Mol Biol. ;705:175-194.

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk