Insertion of foreign random sequences of 120 amino acid residues into an active enzyme.

Abstract:

:Random sequences of 120-130 amino acid residues were inserted into a surface loop region of Escherichia coli RNase HI. This library was screened and about 10% of the clones were found to retain RNase H activity. Subsequent random mutagenesis led to an increase in RNase H activity and solubility of the protein. The inserted regions were found not to contribute to the secondary structure of the mutant protein. The high frequency of insertion of flexible sequences and the increase in the protein's function by further mutagenesis simulate one of the events in protein evolution.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Doi N,Itaya M,Yomo T,Tokura S,Yanagawa H

doi

10.1016/s0014-5793(96)01522-0

subject

Has Abstract

pub_date

1997-02-03 00:00:00

pages

177-80

issue

2-3

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(96)01522-0

journal_volume

402

pub_type

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