Extraction and bioinformatics analysis of Chlamydia trachomatis LpxA
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ORIGINAL PAPER
Extraction and bioinformatics analysis of Chlamydia trachomatis LpxA De-Kun Li . Huan-Huan Feng . Ying-Tao Mu Fang Yang
. Jin-Qiang Yu .
Received: 6 April 2020 / Accepted: 7 October 2020 Ó Springer Nature B.V. 2020
Abstract 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 ahelix, extended strand, b-turn and random coil accounted for 19.6%, 32.8%, 11.4% and 36%,
De-Kun Li and Huan-Huan Feng have contributed equally to this work. D.-K. Li J.-Q. Yu F. Yang Department of Ophthalmology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China H.-H. Feng Xiangyang No.1 People’s Hospital, Hubei University of Medicine, Xiangyang, Hubei, China Y.-T. Mu (&) Department of TCM, Renmin Hospital, Hubei University of Medicine, No. 39 Middle Chaoyang Road, Shiyan 442000, Hubei, China e-mail: [email protected]
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 Chlamydia trachomatis LpxA protein Gene cloning Bioinformatics
Introduction Trachoma is an infectious keratoconjunctivitis caused by Chlamydia trachomatis (Ct), which was very common in China before 1950. And now there are still many trachoma patients in some backward areas of our country. In some underdeveloped areas of Asia, Africa and Latin America, the incidence is still very high [1]. According to the World Health Organization (WHO) survey, it was estimated that more than 8 million people around the world suffered from visual impairment due to trachoma infection. And about 5.5 million people suffered from severe vision loss, or even blindness [2]. Trachoma is still a very important public health problem so far. Ct, as gram-negative bacteria, is an obligate intracellular pathogen. It was first isolated from trachoma patients by Chinese microbiologist Feifan Tang and cultured in vitro. Ct is the primary cause of infectious
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blindness and one of the most widely spread pathogens of sexually transmitted diseases. It was found that serotypes A, B and C were easy to cause ocular infection and infectious trachoma [2–4]. Ct has a unique two-phase development cycle. Now it has been confirmed that Ct has many virulence factors, such as plasmid protein, heat shock pro
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