Cloning of Endoglucanase I Gene from Trichoderma longi-brachiatum and Its Expression in Pichia pastoris
- VernacularTitle:长梗木霉内切葡聚糖酶I基因的克隆及其在毕赤酵母中的表达
- Author:
Hai-Ying LIU
;
Juan WANG
;
Zheng-Yu SHU
;
Song-Gang WU
;
Jian-Zhong HUANG
;
- Publication Type:Journal Article
- Keywords:
Molecular marker, Cellulase endoglucanase I, Pichia pastoris, Expression
- From:
Microbiology
1992;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
A cellulase high-yield strain was identified and named as Trichoderma longibrachiatum SSL by ITS sequence identification. The endoglucanase1 gene (eg1) encoding endo-l,4-?-D-glucanase I was ampli-fied by RT-PCR method, which including 1386 bp and encoding 461 amino acid. Sequence analysis showed that: This gene has a more 90% homology with the T. longibrachiatum eg1 gene. The eg1 gene encoding the mature peptide was inserted into the Pichia pastoris expression vector pPIC9K, which resulted in construc-tion of the recombinant expression plasmid, pPIC9k-eg1. The pPIC9k-eg1 was then introduced into the host Pichia pastoris GS115. After the induction of methanol, extracellular recombinant endoglucanase I from the supernatant of the recombinant Pichia pastoris strain reached 73 U/mL. A clear strengthening of the protein bands, whose molecular weight is about 58 kD, appeared in the SDS-PAGE.
