Abstract: Objective To obtain human enolase - α( ENO1) recombinant protein by prokaryotic expression technology, and carry out bioinformatics analysis of the expressed product. Methods According to the ENO1 gene sequence (cDNA clone BC015641 MGC:23319 IMAGE:4643088) contained in GenBank, the specific primers of the gene fragment were designed. The ENO1 sequence was amplified by PCR and the clone of the gene was constructed by using the cDNA of ENO1 as a template. The vector pEASY-T1 / ENO1 and the expression vector pET-32а(+)/ENO1 were induced to express the recombinant protein ENO1 by IPTG. The bioinformatics related software (ProtParam, ProtScale, SignalP 4.1 server, Signal-3L, TMpred, DAS, NetPhos 2.0 Server) was used to analyze the physicochemical properties, signal peptides, transmembrane domains and phosphorylation sites of the recombinant protein ENO1. Results The recombinant plasmid pEASY-T1 / ENO1 and the expression vector pET-32а(+)/ENO1 were successfully constructed. The coincidence rate of the cloned plasmids and the ENO1 gene sequence in GenBank was 100%. The obtained rENO1 was about 67 ku. The ENO1 fragment consisted of 434 amino acids with a relative molecular mass of 47168.96 and a theoretical pI of 7.01, which is a stable hydrophilic protein. Conclusion Human α-dense alcoholase can be obtained by prokaryotic expression technology. The obtained fusion protein fragment is a stable hydrophilic extracellular protein, which lays a foundation for rapid early diagnosis of liver disease related markers.

Key words: Enolase-α, prokaryotic expression, bioinformatics