免疫共沉淀筛选牛卵泡CART相互作用蛋白的研究

doi:10.11843/j.issn.0366-6964.2015.12.007

Abstract

Abstract:

The study is focused on screening the proteins interacting with cocaine-and amphetamine-regulated transcript(CART) during follicle development in bovine.Four 10-month-old healthy cows，treated for estrus synchronization，were monitored by B-ultrasonic machine to find the ovarian follicles with the diameter of 8-12 mm，which were collected followed by cows being slaughtered.The granulosa cells(GCs) were isolated from those ovarian follicles and then total protein was extracted.CART and the binding protein were deposited by primary antibodies(Rabbit anti CART) using Protein G-Agarose magnetic beads，followed by salt and miscellaneous protein being removed by washing repeatedly.Protein complexes was analyzed by LC-ESI-Q-TOF mass spectrometry(MS) and alignment followed by being eluted and digested by trypsin.Results showed that 111 proteins were identified，in which 81 proteins were functional in conformity with the interaction network modules by Cytoscape V3.1.0 analysis.For the function enrichment analysis，those 81 proteins were classified into 34 groups in 3 categories：molecular function(11.73%)，biological processes(46.93%)，cell component(41.34%).The correlation coefficient of the interaction between A2M，VIM and CART was maximum.It was also found CART was negatively regulated by NALP1，SERPINH1 and PDIA6，which suggested that they might participate in regulating follicle development in bovine.

CLC Number:

S823.2

LI Peng-fei，MENG Jin-zhu，LIU Yan，HUANG Yang，CHEN Jian-wei，JIANG Xiao-long，CAO Xia，YAO Xiao-lei，ZHAO Miao-miao，Lü Li-hua. Screening Proteins Interacting with Cocaine-and Amphetamine-Regulated Transcript in Bovine Follicles by Co-immunoprecipitation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(12): 2169-2175.

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Screening Proteins Interacting with Cocaine-and Amphetamine-Regulated Transcript in Bovine Follicles by Co-immunoprecipitation

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