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Dual Expression of Mouse and Rat VRL-1 in the Dorsal Root Ganglion Derived Cell Line F-11 and Biochemical Analysis of VRL-1 After Heterologous Expression

Tác giả: Ricarda Jahnel, Olaf Bender, Lisa M. Münter, Mathias Dreger, Clemens Gillen, Ferdinand Hucho

Lĩnh vực: Neurochemistry, Biochemistry

Nội dung tài liệu: This study investigates the dual expression of mouse and rat VRL-1 (TRPV2) in the F-11 cell line, a hybridoma derived from mouse neuroblastoma and rat dorsal root ganglion. The research details the cloning and biochemical analysis of VRL-1, including its glycosylation and cellular localization, comparing endogenous and heterologously expressed forms. The findings suggest that F-11 cells offer a valuable experimental system for studying VRL-1 function, with the important consideration that both mouse and rat VRL-1 proteins are present.

Mục lục chi tiết:

  • Introduction
  • Keywords
  • Experimental procedures
  • Reverse transcription of rat brain, F-11 and N18TG2 total RNA
  • VRL-1 specific PCR
  • Single cell RT-PCR of F-11 cells
  • Non-denaturing PAGE
  • Cloning of rVRL-1
  • Cell culture and transfection of F-11 cells
  • Preparation of F-11 cell homogenate and particulate
  • Western blot analysis
  • Deglycosylation with N-glycanase and endoglycosidase H
  • Results
  • VRL-1 is expressed in F-11 cells
  • Mouse and Rat VRL-1 are both expressed in F-11 cells
  • Localization of heterologously expressed vs. endogenous VRL-1 in F-11
  • rVRL-1 is a glycoprotein
  • Discussion
  • Acknowledgements
  • References