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来自中国福建省的三条CRF01_AE全长1型艾滋病毒序列的基因特征分析。

Genetic characterization of three CRF01_AE full-length HIV type 1 sequences from Fujian Province, China.

作者信息

Huang Hai-long, Yan Ping-ping, Zheng Jian, Wu Shou-li, Cheng Ge, Lin Xun, Zheng Wu-xiong, Xie Mei-rong, Zhang Jian-ming, Yan Yan-sheng

机构信息

Fujian Medical University, Fujian 350001, China.

出版信息

Chin Med J (Engl). 2006 Oct 5;119(19):1622-8.

Abstract

BACKGROUND

One of the major characteristics of the human immunodeficiency virus type 1 (HIV-1) is its unusually high degree of genetic variability, which involves in genetic diagnosis, subtyping, vaccine design, and epidemiology. HIV-1 CRF01_AE is a main prevalent HIV-1 recombinant strain in China. In this study, three full-length CRF01_AE genomes from Fujian Province, China were cloned, sequenced, and analyzed; and the further genetic diversity defining and epidemiologic analysis were carried out.

METHODS

Proviral DNA was extracted from non-cultured peripheral blood mononuclear cells, the near full-length HIV-1 genome was amplified and the PCR products were cloned into pCR-XL-TOPO vector and sequenced. 5'-long terminal repeat (LTR) and 3'-LTRs were amplified by additional independent PCR and cloned into pMD18T vector. Gene-based phylogenic tree was constructed and genetic distances were calculated by MEGA 3.1. Simplot was used for Bootscan analysis.

RESULTS

The phylogeny and genetic distance analysis of the three near full-length sequences confirmed that these three samples clustered with CRF01_AE isolates, more close to Thailand CRF01_AE strain CM240, and were distantly related to African CRF01_AE strain 90CF402. Analysis of their genomic organization revealed the presence of nine potential open reading frames. There were no major deletions, rearrangements, or insertions in the three sequences, but an in-frame stop codon was found in tat gene of Fj051. LTRs of the three sequences contained a few nucleotides mutation. We did not find new mosaic recombinant in the three sequences. The V3 motif was GPGQ in all the three sequences, and there were only few amino acids differences in all three V3 loop sequences.

CONCLUSION

This report reveals the background of the three full-length CRF01_AE genomes, the most dominantly circulating HIV-1 strain in Fujian Province, China. The work is essential for the design and development of an effective AIDS vaccine for the region.

摘要

背景

人类免疫缺陷病毒1型(HIV-1)的主要特征之一是其异常高的基因变异性,这涉及基因诊断、亚型分析、疫苗设计和流行病学研究。HIV-1 CRF01_AE是中国主要流行的HIV-1重组毒株。本研究对来自中国福建省的三个全长CRF01_AE基因组进行了克隆、测序和分析,并进行了进一步的基因多样性界定和流行病学分析。

方法

从未培养的外周血单核细胞中提取前病毒DNA,扩增近乎全长的HIV-1基因组,将PCR产物克隆到pCR-XL-TOPO载体中并测序。通过额外的独立PCR扩增5'长末端重复序列(LTR)和3'-LTR,并克隆到pMD18T载体中。构建基于基因的系统发育树,并使用MEGA 3.1计算遗传距离。使用Simplot进行Bootscan分析。

结果

对三个近乎全长序列的系统发育和遗传距离分析证实,这三个样本与CRF01_AE分离株聚类,更接近泰国CRF01_AE毒株CM240,与非洲CRF01_AE毒株90CF402亲缘关系较远。对其基因组结构分析显示存在九个潜在的开放阅读框。这三个序列中没有重大缺失、重排或插入,但在Fj051的tat基因中发现了一个框内终止密码子。这三个序列的LTRs包含一些核苷酸突变。在这三个序列中未发现新的嵌合重组。所有三个序列中的V3基序均为GPGQ,并且所有三个V3环序列中只有少数氨基酸差异。

结论

本报告揭示了中国福建省最主要流行的HIV-1毒株——三个全长CRF01_AE基因组的背景。该工作对于该地区有效艾滋病疫苗的设计和开发至关重要。

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