[Evaluation of construction and expression shiga toxin2 A-luteinizing hormone releasing hormone recombinant toxin].
OBJECTIVE
The purpose is to construct shiga toxin2A-luteinizing hormone releasing hormone (Stx2A-LHRH) recombinant toxin which can kill cancer cells specifically and try to get a new target-binding drug.


METHODS
The fragment of Stx2A-LHRH DNA amplified by polymerase chain reaction (PCR) were cloned into plasmid pET-20b(+) vector. The recombinant plasmid pET-Stx2A-LHRH was constructed successfully and identified by endonucleases digestion and sequencing analysis then it was transformed to Escherichia coli BL21 (DE3) and expressed under the induction of isopropyl-beta-D-thiogalactopyranoside.


RESULTS
The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel thin layer scanning proved a special band of a molecular weight of about 38,000, accounting for 10.32% of total amount of the supernatant protein. Stx2A-LHRH recombinant toxin can kill HeLa cells clearly.


CONCLUSIONS
Stx2A-LHRH recombinant toxin may become a choice of target-binding drug in the future.
Yue
Ying
Y
Department of Obstetrics and Gynecology, First Hospital Jilin University, Changchun 130021, China.
Yu
Xiao-wei
XW
Yue
Yu-huan
YH
Feng
Shu-zhang
SZ
Zhu
Ping
P
Li
He-lian
HL
Yue
Jie
J
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D022641:Shiga Toxin 2 / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D022641:Shiga Toxin 2; D007987:Gonadotropin-Releasing Hormone