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小鼠中枢神经系统中一种新的髓鞘碱性蛋白转录变体。

A novel myelin basic protein transcript variant in the murine central nervous system.

作者信息

Valdivia Anddre Osmar, Farr Valentina, Bhattacharya Sanjoy K

机构信息

Bascom Palmer Eye Institute, University of Miami, Miami, FL, USA.

Neuroscience Graduate Program, University of Miami, Miami, FL, USA.

出版信息

Mol Biol Rep. 2019 Apr;46(2):2547-2553. doi: 10.1007/s11033-019-04635-8. Epub 2019 Feb 12.

Abstract

Myelin basic protein is a multifunctional protein whose primary role is to adhere membranes of the myelin sheath. There are various isoforms that have been identified, 6 distinct isoforms in human and 13 distinct isoforms in mice. These distinct isoforms are the product of alternative splicing of a single gene. The present study sought out to identify the different isoforms found in the murine central nervous system. Neuronal tissue (brain) from five different C57BL6/J mice at 2 months of age was harvested and used for mRNA extraction. mRNA was reversed transcribed to cDNA and transcripts were detected through PCR amplification and DNA agarose gel separation. Primers for exon 1, exon 5b and exon 11 of the myelin basic protein gene were used to capture all the possible transcripts that are naturally found in the murine central nervous system. Unknown transcript was sequenced at Genewiz facilities (South Plainfield, NJ) and mass spectrometry protein sequence analysis demonstrated the presence of a novel myelin basic protein transcript variant. We identified a novel transcript variant of myelin basic protein. This novel transcript variant corresponds to a myelin basic protein of 32.5 kDa which has not been previously reported. This novel transcript variant presents relevant clinical significance to various demyelinating diseases due to its contribution to the understanding of the natural state of the murine central nervous system.

摘要

髓鞘碱性蛋白是一种多功能蛋白,其主要作用是黏附髓鞘的膜。已鉴定出多种异构体,人类有6种不同的异构体,小鼠有13种不同的异构体。这些不同的异构体是单个基因可变剪接的产物。本研究旨在鉴定在小鼠中枢神经系统中发现的不同异构体。采集了5只2月龄C57BL6/J小鼠的神经元组织(脑)并用于提取mRNA。mRNA逆转录为cDNA,并通过PCR扩增和DNA琼脂糖凝胶分离检测转录本。使用髓鞘碱性蛋白基因外显子1、外显子5b和外显子11的引物来捕获在小鼠中枢神经系统中自然存在的所有可能的转录本。未知转录本在Genewiz公司(新泽西州南普莱恩菲尔德)进行测序,质谱蛋白质序列分析表明存在一种新的髓鞘碱性蛋白转录本变体。我们鉴定出一种新的髓鞘碱性蛋白转录本变体。这种新的转录本变体对应于一种32.5 kDa的髓鞘碱性蛋白,此前尚未见报道。这种新的转录本变体因其有助于理解小鼠中枢神经系统的自然状态,对各种脱髓鞘疾病具有重要的临床意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc76/6472899/2e34e725af1c/nihms-1521461-f0001.jpg

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