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A new kind of HSP90 and HSP70 chaperone protein TTC36 localization and function in human retinal pigment epithelial cells and gene LimsA clone analysis

Author: ChenZuo
Tutor: LiuQingHuai
School: Nanjing Medical University
Course: Ophthalmology
Keywords: Heat shock protein 90 Heat shock protein 70 TTC36 ARPE-19 Proliferative vitreoretinopathy Lims gene Gene Expression Gene shear
CLC: R774.1
Type: Master's thesis
Year: 2009
Downloads: 17
Quote: 0
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Abstract


Objective: To study the heat shock protein HSP90 and HSP70 chaperone protein TTC36 subcellular localization and function in human retinal pigment epithelial (RPE) cells in ARPE-19 , to further explore TTC36 and of HSP90 and HSP70 in the RPE cells proliferation and apoptosis, and RPE cell proliferation associated lesions such as proliferative vitreoretinopathy (PVR) occurrence and development process of the mechanism . : Recombinant to build of TTC36 red fluorescence expression vector , with LipofectamineTM 2000 the plp-RFP-TTC36 plp-GFP-HSP90/plp-GFP-HSP70 total transfected RPE cells , laser confocal microscope TTC36 and HSP90/70 co-localization in ARPE-19 cells . ARPE-19 cells transfected TTC36 cell cycle by flow cytometry . The real-time PCR detection of TTC36, HSP90 expression and HSP70mRNA in ARPE-19 cells HSP90 inhibitor 17-AAG treatment level . Results : PCR and sequencing analysis identified successfully build a TTC36 red fluorescent the plp - RFP - TTC36 expression vector ; TTC36 and HSP90, TTC36 and HSP70 are co-localized in the cytoplasm of ARPE-19 cells , and TTC36 transient transfection of promoting ARPE-19 cell proliferation . 17-AAG treatment ARPE-19 cells led to TTC36 mRNA expression decreased . CONCLUSIONS : RPE cells the TTC36 and the heat shock protein of HSP90 HSP70 can interact , therefore TTC36 and HSP90, HSP70 may be involved in the proliferation and apoptosis of RPE cells may be associated with some of RPE cells proliferation and apoptosis associated lesions such as proliferative vitreoretinopathy the occurrence and development . This study TTC36 and heat shock protein of HSP90 , HSP70 function of research laid the foundation , and possible to study the RPE cells proliferation and apoptosis-related eye disease pathogenesis and treatment has opened up a new way . Objective: cloning and analysis of rat shear the Lims gene . Method : using nested RT - PCR , SD rat cDNA template, Lims gene was amplified shear sub- structure into PinPointTM Xa-1T plasmid sequencing . Results: DNA sequencing showed expression of the the the new shear the Lims gene variation Lims A , the variation of shear coding region of 1014bp, encoding 338 amino acids . The conclusion : comparative genomics analysis shows that successfully cloned a new rat the Lims gene the shear sub Lims A lay the foundation for further exploration of the the Lims gene function in proliferative vitreoretinopathy .

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CLC: > Medicine, health > Ophthalmology > Retina and optic nerve diseases > Retinal disease
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