[The expression and localization of p27(KIP1)-GFP fusion gene in hepatocellular carcinoma cell lines].
OBJECTIVE
To study the expression and localization of p27(KIP1)-GFP fusion protein in hepatocellular carcinoma cell line HCC-9204.


METHODS
According to the encoded sequence of human p27(KIP1) gene, stop codon TAA located at 3' terminus of p27(KIP1) gene was mutated into TGT by PCR and fused with GFP gene. Then the p27(KIP1)-GFP cDNA was transfected into HCC-9204 cells. The expression of p27(KIP1)-GFP fusion protein was observed under fluorecence microscope.


RESULTS
DNA sequencing demonstrated that difference between p27(KIP1) gene and that in Genbank was only one nucleoside and the encoded protein sequence is identical. The observation under fluorecence microscope reporced showed that fluorochrome was spread in entire HCC-9204 cells transfected with GFP gene, whereas green fluorescence was seen mainly in the nucleus of HCC-9204 cells transfected with p27(KIP1)-GFP gene.


CONCLUSIONS
The human p27(KIP1)-GFP fusion protein could be expressed in HCC-9204 cells and located in the nucleus, which is similar to the expression of p27(KIP1) gene.
Zheng
Jian-Yong
JY
Department of Gastrointestinal Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Li
Jiang
J
Li
Kai-Zong
KZ
Wang
Wen-Liang
WL
Wang
Wei-Zhong
WZ
D006528:Carcinoma, Hepatocellular; D018797:Cell Cycle Proteins / Q000235:genetics; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms
D018797:Cell Cycle Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27