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Name: Leung Kin Man
University number: 2009109720
Date of experiment: 25-2-2010

BIOL1133: biologic Science Laboratory Course
Experiment 4: Expression of humans secretin receptor concretion protein in E coli.

Aim:
The suggest of the experiment is to express human secretin receptor fusion protein in E coli by conducting SDS-PAGE gel electrophoresis and estimate the molecular size of that protein.

Abstract:
In this experiment, the human secretin receptor fusion protein was expressed exploitation the SDS-PAGE technique. The pre-induced and the sample after IPTG induction was use to extract the human secretin receptor protein by using denaturing buffer zone and elution buffer. The molecular weight of the protein can also be estimated by comparing the bands on the protein ladder.

Introduction:
Massive production of active protein is usually done by recombinant deoxyribonucleic acid technology. E coli. was employ as vector for insertion and expression of fusion protein. E coli. is a germ-negative bacteria with double membrane layer and gamin plasmid.
In this experiment, the N-terminal extracellular domain of the human secretin receptor in E coli. was expressed using IPTG inducible system.



Secretin is the key hormone that prudent for the regulation of gastrointestinal function by regulating the secretion of bicarbonate and enzyme from the exocrine pancreas. This can help to neutralize stomachic acid (hydrochloric acid) and hence maintain an optimum pH care for for intestinal digestion. The action of secretin is mainly controlled by high-affinity membrane receptors distributed in our body and it is tissue-specific. The N-terminal coding region of the extracellular domain of the receptor protein is obtained by conducting polymerase chain reaction (PCR) mutagenesis method. This technique was achieved by using specific DNA primers and DNA strand as a template to start the PCR reaction. The coding sequence was introduced into the bacterial DNA as a carrier of the coding sequence. Upon IPTG...If you want to posture a full essay, order it on our website: Orderessay



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