可卡因苯丙胺调节转录肽相互作用蛋白的筛选

基础医学与临床 ›› 2007, Vol. 27 ›› Issue (1): 25-30.

可卡因苯丙胺调节转录肽相互作用蛋白的筛选

陈娟赵瑛由振东

第二军医大学长征医院神经内科第二军医大学长征医院神经内科第二军医大学神经生物教研室

收稿日期:2005-09-15 修回日期:2006-04-29 出版日期:2007-01-25 发布日期:2007-01-25
通讯作者: 陈娟

Screening of proteins interacting with neuropeptide CART

Received:2005-09-15 Revised:2006-04-29 Online:2007-01-25 Published:2007-01-25

摘要/Abstract

摘要： 摘要：目的寻找可卡因苯丙胺调节转录肽(CART)受体或与其相互作用的蛋白。方法建立大鼠脑cDNA文库，以CART为诱饵，应用细菌双杂交方法筛选与CART相互作用的蛋白。结果成功构建大鼠脑pTRG-cDNA文库及诱饵质粒pBT- CART41-102。文库容量为3.37×106,重组率98.5%。筛选约5.01×106 个共转化克隆，证实为阳性克隆93个，全部DNA测序。经生物信息学方法分析得到stathmin等6个可能与CART相互作用的蛋白，并得到可能与CART相互作用的未知新蛋白，它们与22个已知膜蛋白或受体有短的极相似片段。结论 CART可能与stathmin等6个已知蛋白相互作用，并可能与一些未知蛋白相互作用，这些未知蛋白与22个膜蛋白或受体有短的相似保守结构。

关键词: cDNA文库, CART, 细菌双杂交系统, 蛋白质-蛋白质相互作用

Abstract: Abstract : Objective: The objective was to look　for cocaine-and amphetamine-regulated　transcript peptide(CART) receptors or proteins interacted to CART. Method :A cDNA library from rat brain was constructed. Then the BacterioMatch two-hybrid system was used to screening for proteins interacted with CART by using CART41-102 as bait. Result:The cDNA library and bait pBT- CART41-102 were constructed successfully. The library harvested 3.37×106 individuals, and the reconstruction rate was 98.5%. 5.01x106 cotransformed clones were screened, 93 clones were conformtively identified to interacting with CART and were DNA sequenced. By bioinformation analysis, 6 known proteins interacted to CART were identified. Other unknown interaction proteins that had short exactly similiar motif to 22 membraneproteins or receptors were also identified. Conclusion: CART might interact to proteins about energy balance, cell proliferation and enzymes related to protein processing and degrading .

Key words: cDNA library, CART, bacterial two-hybrid system, protein-protein interaction

陈娟赵瑛由振东. 可卡因苯丙胺调节转录肽相互作用蛋白的筛选[J]. 基础医学与临床, 2007, 27(1): 25-30.

. Screening of proteins interacting with neuropeptide CART[J]. Basic & Clinical Medicine, 2007, 27(1): 25-30.