Expression of human cardiac muscle type fatty acid binding proteins in bacillus coli DH5 alpha and purification

Abstract

The invention relates to the expression and purification of human heart-type fatty acid-binding protein in escherichia coli DH5 alpha, which belongs to the biological technical field, the expression and purification comprises: firstly, preparing an oligonucleotide primer, secondly, amplifying with RT-PCR and determining a sequence, thirdly, constructing recombinant expression plasmids, fourthly, expressing heart-type fatty acid-binding protein through inducing, and fifthly, extracting and purifying the human heart-type fatty acid-binding protein. The expression and the purification of the invention have the beneficial effect that a PL promoter is enabled to enter a complete inhibitory state. H-FABP protein is expressed in PBV220 through utilizing the characteristic. We optimize the temperature and the time in the expression process, bacteria is increased under the temperature of 30DEG C, and high level expression is obtained through inducing for 5 hours under the temperature of 42 DEG C. The successful clone and expression of the human heart-type fatty acid-binding protein lay the foundation of further researching the significance of the heart-type fatty acid-binding protein in the diagnosis and therapeutic effect evaluating of diseases.

Description

technical field

[0001] The invention belongs to the field of biotechnology. Background technique

[0002] 1. Heart-type fatty acid-binding protein (H-FABP)

[0003] (1). The nature and characteristics of FABP

[0004] Fatty Acid Binding Protein (FABP) was first discovered by Ockner et al. in 1972 when studying the regulation of fatty acid absorption in the small intestine of rats, a family of homologous small molecule intracellular proteins found in the intestinal mucosa [1] . The molecular weight is 12-16KD, widely distributed in mammalian small intestine, liver, fat, heart, brain, skeletal muscle and other cells [2] . The discovered FABPs include cardiac muscle type (H-FABP), small intestine type (I-FABP), liver type (L-FABP), adipocyte type (A-FABP), brain cell type (B-FABP), kidney type ( K-FABP), skeletal muscle type (s-FABP), psoriasis-associated type (PA-FABP) and epidermal type (E-FABP) 9 types [1-6] . FABPs are named after their distribution in tissues, and a...

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