Expression and subcellular localization of P9-ZFD protein in patients with myasthenia gravis

Ming Shan Ren, Chuan Zhen Lu, Jian Qiao, Hui Min Ren, Ren Xu, Ren Bao Gan

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Objective: To express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG. Methods: The cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET-24a, and the P9-ZFD recombinant protein was induced via E.coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied. Results: The molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control. Conclusion: P9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Original languageEnglish
Pages (from-to)221-224
Number of pages4
JournalChinese Medical Sciences Journal
Volume19
Issue number3
StatePublished - Sep 2004

Keywords

  • Myasthenia gravis
  • P9-ZFD gene fragment
  • Skeletal muscle

ASJC Scopus subject areas

  • Medicine (all)

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