Cloning and Expression of Trichoderma reesei Cellobiohydrolase I in Pichia pastoris

1999 ◽  
Vol 15 (5) ◽  
pp. 828-833 ◽  
Author(s):  
S. Godbole ◽  
S.R. Decker ◽  
R.A. Nieves ◽  
W.S. Adney ◽  
T.B. Vinzant ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Trichoderma Reesei ◽  
Cloning And Expression ◽  
Cellobiohydrolase I

Cloning and Expression of Full-Length Trichoderma reesei Cellobiohydrolase I cDNAs in Escherichia Coli

1996 ◽  
pp. 389-397
Author(s):  
Robert A. Laymon ◽  
William S. Adney ◽  
Ali Mohagheghi ◽  
Michael E. Himmel ◽  
Steven R. Thomas
Keyword(s):  
Escherichia Coli ◽  
Trichoderma Reesei ◽  
Full Length ◽  
Cloning And Expression ◽  
Cellobiohydrolase I

Cloning and Expression of Recombinant Human Insulin Gene in Pichia pastoris

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Rafid A. Abdulkareem
Keyword(s):  
Pichia Pastoris ◽  
Human Insulin ◽  
Expression Vector ◽  
Expression System ◽  
Insulin Gene ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Major Band ◽  
Human Insulin Gene ◽  
Pichia Pastoris Expression System

The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).

Expression of a secretory β-glucosidase from Trichoderma reesei in Pichia pastoris and its characterization

2011 ◽  
Vol 33 (12) ◽  
pp. 2475-2479 ◽  
Author(s):  
Ping Chen ◽  
Xianyu Fu ◽  
Tzi Bun Ng ◽  
Xiu-Yun Ye
Keyword(s):  
Pichia Pastoris ◽  
Trichoderma Reesei

Cloning and Expression of a Novel Xylanase Gene (Auxyn11D) from Aspergillus usamii E001 in Pichia pastoris

2012 ◽  
Vol 167 (8) ◽  
pp. 2198-2211 ◽  
Author(s):  
Huimin Zhang ◽  
Minchen Wu ◽  
Jianfang Li ◽  
Shujuan Gao ◽  
Yanjun Yang
Keyword(s):  
Pichia Pastoris ◽  
Cloning And Expression ◽  
Xylanase Gene ◽  
Aspergillus Usamii

The diversity of glycosylation of cellobiohydrolase I from Trichoderma reesei determined with mass spectrometry

2019 ◽  
Vol 508 (3) ◽  
pp. 818-824 ◽  
Author(s):  
Mingyu Wang ◽  
Yanan Ma ◽  
Ling Li ◽  
Bianfang Wang ◽  
Xin Wei ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Trichoderma Reesei ◽  
Cellobiohydrolase I

Heterologous expression of codon optimized Trichoderma reesei Cel6A in Pichia pastoris

2016 ◽  
Vol 92 ◽  
pp. 107-116 ◽  
Author(s):  
Fubao Fuelbiol Sun ◽  
Renhui Bai ◽  
Huimin Yang ◽  
Fei Wang ◽  
Jing He ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Heterologous Expression ◽  
Trichoderma Reesei
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