CLONING AND ANALYSIS OF THE CODING SEQUENCE FOR HEPATITIS B VIRUS PRES1 BINDING PROTEIN BY T7 cDNA PHAGE DISPLAY SYSTEM / 解放军医学杂志

ABSTRACT

T7 cDNA phage display method was employed to find the binding protein of PreSl protein. PreSl protein was coated in a 96-well ELISA plate, and then T7 cDNA library phages were bound to the target protein. Phages which did not bind to garget protein were washed away and the binding phages were eluted. Insertions from different clones were sequenced, and the deduced amino acid sequences were analyzed by Vector 6.0 software. Using BLAST software in GenBank, whole length of amino acid sequence of binding protein was obtained. After 4 rounds of biopanning, recombinant T7 phages with binding ablity were amplifed by infection to E. coli. One piece of amino acid sequence was found to be amino terminal of product of glioma tumor suppressor candidate region gene 2 (GLTSCR2). There was a binding domain KxPxKSGxxxL in these clones. T7 cDNA phage display technique can be used bo find the ligand. GLTSCR2 coding protein may be the binding protein to preSl protein of HBV.\;