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中国树鼩(Tupaia belangeri chinensis)中NPC1L1的分子克隆与特性分析

Molecular cloning and characterization of NPC1L1 in the Chinese tree shrew (Tupaia belangeri chinensis).

作者信息

Kui Xiuying, Qiu Dandan, Wang Wenguang, Li Na, Tong Pinfen, Sun Xiaomei, Jin Liangzi, Deng Wei, Dai Jiejie, Lu Caixia

机构信息

Center of Tree Shrew Germplasm Resources, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming, China.

Yunnan Key Laboratory of Vaccine Research and Development On Severe Infectious Diseases, Kunming, China.

出版信息

Mol Biol Rep. 2021 Dec;48(12):7975-7984. doi: 10.1007/s11033-021-06829-5. Epub 2021 Oct 30.

Abstract

BACKGROUND

The Niemann-Pick C1-Like 1 protein, a multi-transmembrane domain molecule, is critical for intestinal cholesterol absorption, and is the entry factor for hepatitis C virus (HCV). The Chinese tree shrew (Tupaia belangeri chinensis) is closer to primates in terms of genetic evolution than rodents. Previous studies indicated that the tree shrew was suitable for HCV research; however, little is known about tree shrew NPC1L1.

METHODS AND RESULTS

TsNPC1L1 cDNA was amplified by rapid amplification of cDNA ends (RACE) technology. The cDNA sequence, its encoded protein structure, and expression profile were analyzed. Results indicated that the tsNPC1L1 mRNA is 4948 bp in length and encodes a 1326 amino acid protein. TsNPC1L1 possesses 84.97% identity in homology to human NPC1L1 which is higher than both mouse (80.37%) and rat (81.80%). The protein structure was also similar to human with 13 conserved transmembrane helices, and a sterol-sensing domain (SSD). Like human NPC1L1, the tsNPC1L1 mRNA transcript is highly expressed in small intestine, but it was also well-expressed in the lung and pancreas of the tree shrew.

CONCLUSION

The homology of tree shrew NPC1L1 was closer to human than that of rodent NPC1L1. The expression of tsNPC1L1 was the highest in small intestine, and was detectable in lung and pancreas. These results may be useful in the study of tsNPC1L1 function in cholesterol absorption and HCV infection.

摘要

背景

尼曼-皮克C1样1蛋白是一种多跨膜结构域分子,对肠道胆固醇吸收至关重要,并且是丙型肝炎病毒(HCV)的进入因子。中华树鼩在基因进化方面比啮齿动物更接近灵长类动物。先前的研究表明树鼩适合用于HCV研究;然而,关于树鼩NPC1L1的了解甚少。

方法与结果

通过cDNA末端快速扩增(RACE)技术扩增树鼩NPC1L1 cDNA。对其cDNA序列、编码的蛋白质结构和表达谱进行了分析。结果表明,树鼩NPC1L1 mRNA长度为4948 bp,编码一个1326个氨基酸的蛋白质。树鼩NPC1L1与人类NPC1L1的同源性为84.97%,高于小鼠(80.37%)和大鼠(81.80%)。其蛋白质结构也与人类相似,有13个保守的跨膜螺旋和一个固醇感应结构域(SSD)。与人类NPC1L1一样,树鼩NPC1L1 mRNA转录本在小肠中高表达,但在树鼩的肺和胰腺中也有良好表达。

结论

树鼩NPC1L1与人类的同源性比啮齿动物NPC1L1与人类的同源性更接近。树鼩NPC1L1在小肠中的表达最高,在肺和胰腺中也可检测到。这些结果可能有助于研究树鼩NPC1L1在胆固醇吸收和HCV感染中的功能。

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