Objective: The aim of this study is to clone the LpxA gene of Chlamydia trachomatis and analyze its biological characteristics.
Methods: Specific primers were designed according to the sequence of Ct LpxA gene. LpxA gene was amplified by PCR and connected to pMD18-T vectors. Positive clones were selected for PCR and DNA sequencing. Finally, bioinformatics software was used to analyze the biological properties of LpxA protein.
Results: The total length of LpxA gene was 840 bp, encoding 280 amino acids. LpxA protein has no signal peptide and was located in bacterial cytoplasm. The prediction of secondary structure showed that the α-helix, extended strand, β-turn and random coil accounted for 19.6%, 32.8%, 11.4% and 36%, respectively. According to the prediction of tertiary structure, three identical LpxA molecules constituted homologous trimers. It was predicted that there were 11 B cell epitopes in LpxA.
Conclusion: Ct Lpxa gene was cloned, and LpxA protein structure and function were predicted.
Keywords: Bioinformatics; Chlamydia trachomatis; Gene cloning; LpxA protein.