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用于乳酸乳球菌亚种嗜热栖热菌MG1363的密码子优化及编码伊朗人乳头瘤病毒16型E7癌基因的质粒构建

Codon Usage Optimization and Construction of Plasmid Encoding Iranian Human Papillomavirus Type 16 E7 Oncogene for Lactococcus Lactis Subsp. Cremoris MG1363.

作者信息

Mohseni Amir Hossein, Razavilar Vadood, Keyvani Hossein, Razavi Mohammad Reza, Khavari Nejad Ramazan Ali

机构信息

Department of Microbiology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, IR Iran. Email:

出版信息

Asian Pac J Cancer Prev. 2017 Mar 1;18(3):783-788. doi: 10.22034/APJCP.2017.18.3.783.

Abstract

HPV 16 intratypic sequence variations has been recognized in association with oncogenic potential diverge and geographic distribution. This study aimed to investigate nucleotide modifications and optimization of HPV 16 E7 regions from Iranian infected women. Cervical biopsies from 79/163 HPV 16 positive cancer patients detected in our study were analyzed by PCR in a couple of cloning of a complete ORF of the E7 gene, and sequencing. The most frequently observed variant was C196T in E7 which led to an amino acid change of R66W. In addition, only one common variant T234G was identified from all specimens, but it did not lead to any amino acid change. We also detected nucleotide variations A86G, and C188T in samples. Among 99 codons in E7 gene, 56 codons were improved for Lactococcus lactis subsp. cremoris MG1363 resulting in a reduced G+C content from 43.1% to 34.0%. Also, the AT%, ENC, and CAI values were 66, 20±1.1, and 1.000 instead of 56.90, 60 ±1.1, and 0.406 respectively. Finally we constructed expression vector pNZ8148 encoding optimized E7 oncoprotein of HPV 16. This study declared for the first time, the genetic variations of HPV 16 E7 in IRAN. We conclude that plasmid pNZ8148-HPV 16-opti E7 can be potential vaccine candidates in the future.

摘要

人乳头瘤病毒16型(HPV 16)的型内序列变异已被认为与致癌潜力差异和地理分布有关。本研究旨在调查伊朗感染女性中HPV 16 E7区域的核苷酸修饰及优化情况。对我们研究中检测出的163例HPV 16阳性癌症患者中的79例进行宫颈活检,通过PCR对E7基因完整开放阅读框进行多次克隆并测序分析。E7中最常观察到的变异是C196T,导致氨基酸R66W改变。此外,在所有样本中仅鉴定出一个常见变异T234G,但它未导致任何氨基酸变化。我们还在样本中检测到核苷酸变异A86G和C188T。在E7基因的99个密码子中,对乳酸乳球菌乳脂亚种MG1363优化了56个密码子,使G + C含量从43.1%降至34.0%。而且,AT%、ENC和CAI值分别为66、20±1.1和1.000,而非之前的56.90、60±1.1和0.406。最后我们构建了编码HPV 16优化E7癌蛋白的表达载体pNZ8148。本研究首次公布了伊朗HPV 16 E7的基因变异情况。我们得出结论,质粒pNZ8148 - HPV 16 - opti E7未来可能成为潜在的疫苗候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb6f/5464500/400cec1c2efc/APJCP-18-783-g001.jpg

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