[PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 gene].
OBJECTIVE
To study the PCR amplification, cloning and protein expression of interferon-inducible transmembrane protein-1 (IFITMP-1) gene.


METHODS
With the cDNA fragment containing IFITMP-1 gene as template, IFITMP-1 gene was amplified using Pfu enzyme by means of PCR. After EcoRI and HindIII digestion, the target gene fragment was linked to pUCm-T plasmid and sequenced. The IFITMP-1 gene was cloned into pET-Trx protein expression plasmid, and the condition for protein expression was optimized.


RESULTS
The length of the PCR product of IFITMP-1 gene-containing cDNA fragment was about 1000 bp. The IFITMP-1 gene was successfully inserted into pUCm-T plasmid with correct sequence and cloning of the IFITMP-1 gene into the pET-Trx protein expression plasmid was achieved. Expression of the fusion protein of pUCm-T plasmid and IFITMP-1 gene was detected after IPTG induction.


CONCLUSIONS
Successful amplification and cloning of the IFITMP-1 gene and its protein expression may facilitate further study of the role of IFITMP-1 gene in colorectal cancer.
Liu
Yu-hu
YH
Department of Digestive Diseases, People's Hospital of Dongguan, Dongguan 523018, China. liuyuhu@126.com
Zhong
Dong
D
Liu
Juan
J
Zhang
Zhen-shu
ZS
Chen
Cun-long
CL
Wu
Jin-bao
JB
Xiao
Bing
B
Guo
Wen-ying
WY
D000943:Antigens, Differentiation; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D000943:Antigens, Differentiation; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C062010:leu-13 antigen; D007372:Interferons