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从阿拉伯单峰驼中克隆 Lin-28 基因及其蛋白的分子特征、生物信息学分析和表达谱研究。

Molecular Characterization, Bioinformatic Analysis, and Expression Profile of Lin-28 Gene and Its Protein from Arabian Camel ().

机构信息

National Center for Stem Cell Technology, King Abdulaziz City for Science and Technology, Riyadh 11442, Saudi Arabia.

National Center for Biotechnology, King Abdulaziz City for Science and Technology, Riyadh 11461, Saudi Arabia.

出版信息

Int J Mol Sci. 2019 May 9;20(9):2291. doi: 10.3390/ijms20092291.

Abstract

Lin-28 is an RNA-binding protein that is known for its role in promoting the pluripotency of stem cells. In the present study, Arabian camel Lin-28 (cLin-28) cDNA was identified and analyzed. Full length cLin-28 mRNA was obtained using the reverse transcription polymerase chain reaction (RT-PCR). It was shown to be 715 bp in length, and the open reading frame (ORF) encoded 205 amino acids. The molecular weight and theoretical isoelectric point (pI) of the cLin-28 protein were predicted to be 22.389 kDa and 8.50, respectively. Results from the bioinformatics analysis revealed that cLin-28 has two main domains: an N-terminal cold-shock domain (CSD) and a C-terminal pair of retroviral-type Cysteine3Histidine (CCHC) zinc fingers. Sequence similarity and phylogenetic analysis showed that the cLin-28 protein is grouped together and . Quantitative real-time PCR (qPCR) analysis showed that cLin-28 mRNA is highly expressed in the lung, heart, liver, and esophageal tissues. Peptide mass fingerprint-mass spectrometry (PMF-MS) analysis of the purified cLin-28 protein confirmed the identity of this protein. Comparing the modeled 3D structure of cLin-28 protein with the available protein 3D structure of the human Lin-28 protein confirmed the presence of CSD and retroviral-type CCHC zinc fingers, and high similarities were noted between the two structures by using super secondary structure prediction.

摘要

Lin-28 是一种 RNA 结合蛋白,其功能是促进干细胞的多能性。本研究鉴定并分析了阿拉伯骆驼 Lin-28(cLin-28)cDNA。使用反转录聚合酶链反应(RT-PCR)获得全长 cLin-28 mRNA。它的长度为 715bp,开放阅读框(ORF)编码 205 个氨基酸。cLin-28 蛋白的分子量和理论等电点(pI)预测分别为 22.389kDa 和 8.50。生物信息学分析结果表明,cLin-28 有两个主要结构域:N 端冷休克结构域(CSD)和 C 端一对逆转录病毒型 Cysteine3Histidine(CCHC)锌指。序列相似性和系统发育分析表明,cLin-28 蛋白与 和 聚为一组。定量实时 PCR(qPCR)分析表明,cLin-28 mRNA 在肺、心、肝和食管组织中高表达。纯化的 cLin-28 蛋白的肽质量指纹图谱-质谱(PMF-MS)分析证实了该蛋白的身份。比较 cLin-28 蛋白的建模 3D 结构与现有的人类 Lin-28 蛋白的 3D 结构,确认了 CSD 和逆转录病毒型 CCHC 锌指的存在,并且通过使用超二级结构预测,两个结构之间存在高度相似性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aabb/6540139/b0c3cf1e2f05/ijms-20-02291-g001.jpg

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