[Construction of a bait plasmid containing HBV PreS1 gene in a yeast two-hybrid system and evaluation of its toxicity and self-activation].
OBJECTIVE
To construct a yeast expression vector of hepatitis B virus (HBV) PreS1 gene using the Sos-recruitment system (SRS), and evaluate the effect of the expression product on the growth of the yeast cells and activation of the reporter gene.


METHODS
The coding sequence of HBV preS1 was amplified by PCR and cloned into the yeast expression plasmid pSos. The recombinant bait plasmid pSos- PreS1 was verified by sequencing before transformation into competent yeast cells. The effects of the expression product on the yeast cell growth and activation of the reporter gene were evaluated.


RESULTS
The yeast expression vector of HBV PreS1 gene was constructed successfully. The recombinant bait plasmid showed no toxic effect on yeast cdc25H cells without a self-activation of the reporter gene.


CONCLUSIONS
The SRS can be used to study the proteins interacting with HBV PreS1 protein and provides a means for obtaining insight into the pathogenic mechanism of HBV.
Department of Infectious Diseases, First Affiliated Hospital, Xi'an Jiaotong University College of Medicine, Xi'an 710061, China.
Xi
X
Zhang
Shu-mei
SM
Lin
Lie-xiu
LX
Wu
Tian-yan
TY
Chen
Feng
F
Ye
Ying-ren
YR
Zhao
Shu-lin
SL
Zhang
D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D011991:Receptors, Virus / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen