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疟蚊 Anopheles stephensi 中纤维蛋白原相关蛋白 1(frep1)的分子特征、表达及计算机分析。

Molecular characterization, expression and in-silico analysis of fibrinogen-related protein 1 (frep1) in malaria vector Anopheles stephensi.

机构信息

Department of Genetics, Maharshi Dayanand University, Rohtak, Haryana, 124001, India.

National Institute of Malaria Research (NIMR), Sector 8, Dwarka, New Delhi, 110077, India.

出版信息

Mol Biol Rep. 2024 Sep 9;51(1):970. doi: 10.1007/s11033-024-09891-x.

Abstract

BACKGROUND

Fibrinogen-related protein 1 (frep1) is a member of the pattern-recognizing receptor family (PRR) which generates an innate immune response after recognizing the pattern associated molecular pattern (PAMP) that occurs on the surface of microorganisms. The main objective of this study is to characterize frep1 and its in-silico analysis in Anopheles stephensi.

METHODS AND RESULT

The DNA was extracted from female Anopheles stephensi. PCR was performed for complete analysis of frep1 using specific primers. The gene sequence of frep1 was identified by Sanger sequencing. The bioinformatics structure analysis approach revealed the presence of 3 exons and 4 introns in the frep1. The sequence of frep1 was submitted to NCBI GeneBank with accession number ON817187.1. Quantitative real-time PCR was performed to analyze frep1 expression. At the developmental stage, frep1 is highly expressed in the L1 stage, egg, and adult female mosquito. In addition, frep1 is highly expressed in the tissue fat body, midgut, and salivary gland. After blood-fed, an upregulation of frep1 at 48 h in the midgut, and downregulation in fat body were observed at different time intervals.

CONCLUSION

The genomic data of frep1 is encoded by 12,443 bp. The frep1 has a significant role in the early metamorphosis. Its expression in fat body and midgut suggests it could be important for fat metabolism and post-blood digestion. The conserved domain could be targeted for vector control. Further study is required to elucidate its function against malaria parasites to confirm its agonist role in malaria transmission.

摘要

背景

纤维蛋白原相关蛋白 1(frep1)是模式识别受体(PRR)家族的成员,在识别微生物表面存在的模式相关分子模式(PAMP)后,会产生先天免疫反应。本研究的主要目的是研究 frep1 及其在斯氏按蚊中的特性。

方法和结果

从雌性斯氏按蚊中提取 DNA。使用特定引物进行 PCR 以完成 frep1 的完整分析。通过 Sanger 测序鉴定 frep1 的基因序列。生物信息学结构分析方法表明 frep1 存在 3 个外显子和 4 个内含子。frep1 序列已提交给 NCBI GeneBank,登录号为 ON817187.1。进行定量实时 PCR 以分析 frep1 的表达。在发育阶段,frep1 在 L1 期、卵和成年雌性蚊子中高度表达。此外,frep1 在组织脂肪体、中肠和唾液腺中高度表达。吸血后,在不同时间点观察到中肠中 frep1 在 48 小时时上调,脂肪体中下调。

结论

frep1 的基因组数据由 12443bp 编码。frep1 在早期变态发育中具有重要作用。其在脂肪体和中肠中的表达表明它可能对脂肪代谢和吸血后消化很重要。保守结构域可以作为载体控制的靶标。需要进一步研究以阐明其对疟原虫的作用,以确认其在疟疾传播中的激动剂作用。

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