Transformation of pPICZaA-E2 to Pichia pastoris X-33 and Mut+ phenotype analysis

2021 ◽  
Author(s):  
F. Shabihah ◽  
S. Pambudi ◽  
F. A. Sitepu ◽  
C. Ikhsan ◽  
B. Yohan ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Phenotype Analysis

Molecular profiling and antimicrobial susceptibility of Escherichia coli O157:H7 isolated in Bulgaria

2020 ◽  
Vol 23 (3) ◽  
pp. 310-318
Author(s):  
K. Koev ◽  
T. Stoyanchev ◽  
G. Zhelev ◽  
P. Marutsov ◽  
K. Gospodinova ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Escherichia Coli O157 ◽  
Acid Sensitivity ◽  
Lower Extent ◽  
Phenotype Analysis ◽  
Anal Swab ◽  
Amoxicillin Clavulanic Acid ◽  
Primary Identification ◽  
Stx1 Gene ◽  
Cattle Farms

The purpose of this study was to detect the presence of shiga-toxin producing Escherichia coli (STEC) in faeces of healthy dairy cattle and to determine the sensitivity of isolates to several anti­microbial drugs. A total of 1,104 anal swab samples originating from 28 cattle farms were examined. After the primary identification, 30 strains were found to belong to serogroup О157. By means of conventional multiplex PCR, isolates were screened for presence of resistance genes stx1, stx2 and eaeА. Twenty-nine strains possesses amplicons with a size corresponding to genes stx2 and eaeA, one had amplicons also for the stx1 gene and one lacked amplicons of all three genes. Twenty-eight strains demonstrated amplicons equivalent to gene H7. The results from phenotype analysis of resistance showed preserved sensitivity to ceftriaxone, ceftazidime, cefotaxime, cephalothin, streptomycin, gentamicin, tetracycline, enrofloxacin and combinations sulfamethoxazole/trimethoprim and amoxicillin/clavulanic acid. Sensitivity to ampicillin was relatively preserved, although at a lower extent.

Optimization of fermentation conditions for the production of recombinant hpdgf-bb in Pichia pastoris

2015 ◽  
Vol 37 (1se) ◽  
Author(s):  
Duong Long Duy ◽  
Pham Minh Vu ◽  
Nguyen Tri Nhan ◽  
Tran Linh Thuoc ◽  
Dang Thi Phuong Thao
Keyword(s):  
Pichia Pastoris ◽  
Fermentation Conditions ◽  
Optimization Of Fermentation

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).

SECRETED EXPRESSION OF LITOPENAEUS VANNAMEI LYSOZYME GENE IN PICHIA PASTORIS AND ITS BIOACTIVITY DETECTION

2010 ◽  
Vol 36 (6) ◽  
pp. 1091-1096
Author(s):  
Shu-Guang BIAN ◽  
Hua-Xin CHEN ◽  
Peng JIANG ◽  
Hai-Bo ZHANG ◽  
Zhao-Pu LIU ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Litopenaeus Vannamei ◽  
Lysozyme Gene ◽  
Secreted Expression

Expression and Characteristics of Phytases from Obesumbacterium proteus in Pichia pastoris Yeast

2018 ◽  
Vol 34 (4) ◽  
pp. 18-25 ◽  
Author(s):  
T.L. Gordeeva ◽  
◽  
L.N. Borshchevskaya ◽  
A.N. Kalinina ◽  
S.P. Sineoky ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Pichia Pastoris Yeast

High-Level Secretory Expression and Purification of Recombinant Human Interleukin 1 Beta in Pichia pastoris

2016 ◽  
Vol 23 (8) ◽  
pp. 763-769 ◽  
Author(s):  
Pengfei Li ◽  
Ganggang Yang ◽  
Xiaofang Geng ◽  
Jinbao Shi ◽  
Bin Li ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Interleukin 1 ◽  
Interleukin 1 Beta ◽  
Expression And Purification ◽  
Secretory Expression ◽  
High Level
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