[The construction of yeast two-hybrid method in the protein-interactions and its significance in hepatic metastasis of colorectal carcinoma].
OBJECTIVE
To construct the yeast two-hybrid system, and screen the proteins which interact with FasL, and investigate the relationship of FasL and hepatic metastasis of colorectal carcinoma.


METHODS
We have cloned the FasL gene into the pGBKT7 vector as the bait, then screened the fetal liver cDNA library, and have got a series of specific proteins that interact with FasL protein. Using the bioinformatics, we analyzed the interacting proteins in the mechanism of hepatic metastasis of colorectal carcinoma.


RESULTS
We have screened several proteins that interaction with FasL protein, including metallothionein 1K, 1G, 2A, cathepsin B, fatty acid synthase, interferon alpha-inducible protein 27, phospholipid scramblase, Ser/Thr-like kinase, anchor attachment protein, fibulin-5.


CONCLUSIONS
We have successfully constructed the yeast two-hybrid system, and preliminary identified that the interaction between FasL, metallothionein, cathepsin and anchor attachment protein is radically related to the hepatic metastasis of colorectal carcinoma.
Department of General Surgery, Beijing Military Area General Hospital, Beijing 100700, China.
Fu-Yi
FY
Zuo
Shi-Yong
SY
Li
Ping
P
An
Bo
B
Yu
Hui-Yun
HY
Cai
D002401:Cathepsin B / Q000378:metabolism; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms / Q000737:chemistry* / Q000473:pathology; D053222:Fas Ligand Protein; D015723:Gene Library; D006801:Humans; D066298:In Vitro Techniques; D008113:Liver Neoplasms / Q000737:chemistry* / Q000556:secondary; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008668:Metallothionein / Q000378:metabolism; D011485:Protein Binding; D048069:Tumor Necrosis Factors / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics*
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D048069:Tumor Necrosis Factors; D008668:Metallothionein; D002401:Cathepsin B