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Molecular Cloning, and Characterization of an Adenylyl Cyclase-Associated Protein from Gossypium arboreum L.
Authors:WANG Sheng  ZHAO Guo-hong  JIA Yin-hua  DU Xiong-ming
Institution:Cotton Germplasm Division, Cotton Research Institute, Chinese Academy of Agricultural Sciences, Anyang 455000, P.R.China
Abstract:The aim of this study was to clone CAP (adenylyl cyclase-associated protein) gene from Gossypium arboreum L. and develop a platform for expressing and purifying CAP protein, which is a base for the construction and function researches of CAP. In this work, a CAP homolog from cotton (DPL971) ovule was identified and cloned. And the cDNA sequence consisted of an open reading frame of 1416 nucleotides encoding a protein of 471 amino acid residues with a calculated molecular weight of 50.6 kDa. To gain insight on the CAP role in cotton fiber development, the cloned CAP cDNA was expressed. A significant higher yield pure protein was obtained with the chromatographic method. Further experiments showed that the purified protein can bind with the actin in vitro indicating that the recombinant cotton CAP is functional. The procedure described here produced high yield pure protein through one chromatographic step, suitable for further structure-function studies.
Keywords:adenylyl cyclase-associated protein  CAP  cotton fiber  protein expression  protein purification  Gossypium arboreum L
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