METHOD FOR CONSTRUCTION OF A LIBRARY IN HANSENULA POLYMORPHA USING HARS36 SEQUENCE AND THE LIBRARY CONSTRUCTED BY THE SAME

摘要

A method for construction of a library in Hansenula polymorpha using a HARS36 sequence and the library constructed by the same method are provided, thereby constructing a useful gene library having high transformation efficiency and uniformed copy number in Hansenula polymorpha without Escherichia coli. The method for construction of the library in Hansenula polymorpha using the HARS36 sequence comprises the steps of: (1) preparing an insertion gene fragment comprising the HARS36 sequence having the nucleotide sequence of SEQ ID NO:1 of which total or portion sequence is exposed to the 5'- or 3'-terminal of the insertion gene fragment and a target expression gene; (2) a receiving gene fragment comprising a selection marker gene, a promoter for the selection marker gene, and the HARS36 sequence having the nucleotide sequence of SEQ ID NO:1 of which total or portion sequence is exposed to the 5'- or 3'-terminal of the receiving gene fragment; (3) transforming Hansenula polymorpha with both gene fragments of steps (1) and (2) to prepare a transformant; and (4) culturing the transformant.