DNA Length Heterogeneity Adjacent to the Human Insulin Gene in Diabetes mellitus

2015 ◽  
pp. 200-211
Author(s):  
G. A. Hitman ◽  
L. G. Williams ◽  
N. I. Jowett ◽  
K. M. Spencer ◽  
D. J. Galton
Keyword(s):  
Diabetes Mellitus ◽  
Human Insulin ◽  
Insulin Gene ◽  
Human Insulin Gene

scholarly journals Seven mutations in the human insulin gene linked to permanent neonatal/infancy-onset diabetes mellitus

2008 ◽  
Author(s):  
Carlo Colombo ◽  
Ottavia Porzio ◽  
Ming Liu ◽  
Ornella Massa ◽  
Mario Vasta ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Human Insulin ◽  
Insulin Gene ◽  
Onset Diabetes ◽  
Human Insulin Gene

Diabetes mellitus, atherosclerosis, and the 5′ flanking polymorphism of the human insulin gene

1986 ◽  
Vol 9 (S1) ◽  
pp. 98-110 ◽  
Author(s):  
T. Mandrup-Poulsen ◽  
D. Owerbach ◽  
J. Nerup ◽  
K. Johansen ◽  
A. Tybjærg Hansen
Keyword(s):  
Diabetes Mellitus ◽  
Human Insulin ◽  
Insulin Gene ◽  
Human Insulin Gene

The Human Insulin Gene and Diabetes Mellitus

2009 ◽  
Vol 87 (01) ◽  
pp. 89-103 ◽  
Author(s):  
Carola Neumer ◽  
Rowena Brandt ◽  
H. Zühlke
Keyword(s):  
Diabetes Mellitus ◽  
Human Insulin ◽  
Insulin Gene ◽  
Human Insulin Gene

Lack of association of the polymorphic locus in the 5'-flanking region of the human insulin gene and diabetes in American blacks

Diabetes ◽  
1985 ◽  
Vol 34 (5) ◽  
pp. 433-439 ◽  
Author(s):  
S. Elbein ◽  
P. Rotwein ◽  
M. A. Permutt ◽  
G. I. Bell ◽  
N. Sanz ◽  
...  
Keyword(s):  
Human Insulin ◽  
Polymorphic Locus ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Flanking Region ◽  
American Blacks

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).

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