Cloning and Expression of Biopharmaceuticals in CHO.K1 and BHK-21 Cells

2005 ◽  
pp. 489-492
Author(s):  
G.F. Asensi ◽  
J.M. Cavalcanti ◽  
P.M. Sousa ◽  
E.M. Del Aguila ◽  
J.T. Silva ◽  
...  
Keyword(s):  
Cloning And Expression

cDNA cloning and expression of a gamma-aminobutyric acidA receptor epsilon-subunit in rat brain

2000 ◽  
Vol 12 (12) ◽  
pp. 4318-4330 ◽  
Author(s):  
Nathalie Moragues ◽  
Philippe Ciofi ◽  
Pierrette Lafon ◽  
Marie-Francoise Odessa ◽  
Gerard Tramu ◽  
...  
Keyword(s):  
Rat Brain ◽  
Cdna Cloning ◽  
Cloning And Expression ◽  
Epsilon Subunit

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).

Cloning and expression analysis of ZFP207 encoding a TFⅢA-type zinc finger protein in rice

2010 ◽  
Vol 32 (4) ◽  
pp. 387-392 ◽  
Author(s):  
Fan-Jun MENG ◽  
Ji HUANG ◽  
Yong-Mei BAO ◽  
Yan JIANG ◽  
Hong-Sheng ZHANG
Keyword(s):  
Zinc Finger ◽  
Expression Analysis ◽  
Zinc Finger Protein ◽  
Cloning And Expression ◽  
Finger Protein

Cloning and Expression Analysis of cMDH Gene Related to Cytoplasmic Male Sterile Wheat with Aegilops kotschyi Cytoplasm

2009 ◽  
Vol 35 (9) ◽  
pp. 1620-1627 ◽  
Author(s):  
Long-Yu ZHANG ◽  
Hong-Xia LI ◽  
Gai-Sheng ZHANG ◽  
Jun-Sheng WANG ◽  
Yan-Fen HAN ◽  
...  
Keyword(s):  
Expression Analysis ◽  
Cloning And Expression ◽  
Male Sterile ◽  
Cytoplasmic Male Sterile

Cloning and Expression Analysis of Galactosyltransferase Gene GhGalT1 Promoter in Cotton

2018 ◽  
Vol 44 (2) ◽  
pp. 218
Author(s):  
Li-Xia QIN ◽  
Jing LI ◽  
Huan-Yang ZHANG ◽  
Sheng LI ◽  
Meng-Jie ZHU ◽  
...  
Keyword(s):  
Expression Analysis ◽  
Cloning And Expression

Cloning and Expression Analysis ofCIPKGene in Sugarcane

2015 ◽  
Vol 41 (3) ◽  
pp. 499 ◽  
Author(s):  
Long HUANG ◽  
Wei-Hua SU ◽  
Yu-Ye ZHANG ◽  
Ning HUANG ◽  
Hui LING ◽  
...  
Keyword(s):  
Expression Analysis ◽  
Cloning And Expression
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