[The construction of PQE-ARL-1 recombinant expression plasmid and the preparation and purification of ARL-1 protein].
OBJECTIVE
To further understand the function and biological activity of aldose reductase like-1 (ARL-1), and to provide the basis for the preparation of the specific antibody against ARL-1.


METHODS
The ARL-1 cDNA was cloned into procaryotic expression vector PQE-30 and the recombinant ARL-1 expressed in E.coli M(15); then Ni(+)-NTA agarose columns were used to purify the ARL-1 protein.


RESULTS
With the digestion of the enzymes, we identified that the ARL-1 gene was inserted into the procaryotic expression vector PQE-30. The expression products of PQE-ARL-1 (human recombinant ARL-1) showed a single protein band on SDS-PAGE. The molecular weight of ARL-1 was approximately 37.7 x 10(3) and the expression level was about 25% of the total bacterial protein. The concentration of ARL-1 protein was about 100mg/L.


CONCLUSIONS
The construction of the recombinant plasmid PQE-ARL-1 and the preparation of the protein ARL-1 have laid a solid foundation for further studying the function of ARL-1 and preparing the specific antibody against ARL-1.
Zheng
C
C
The Center of Molecular Biology, Hengyang Medical College, Hengyang 421001, China.
Wan
L
L
Wu
M
M
Cao
D
D
Yang
Y
Y
D000449:Aldehyde Reductase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D000074408:Aldo-Keto Reductases; D008970:Molecular Weight; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis*
D011994:Recombinant Proteins; C120471:AKR1B10 protein, human; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase