棉 花 学 报        Cotton Science      2009，21(6)：520-523

Expression of a GAI Gene of Cotton ( Gossypium hirsutum ) and Preparation of Its Antibody
ZHENG Yin-ying^1，2，3，CUI Bai-ming^2，3，ZHU Jian-bo³，LIAO Wen-bin²，ZHANG Gen-fa⁴，PENG Ming^1，2，3*
(1.Hainan University, Haikou 570228, China; 2.Institute of Tropical Bioscience and Biotechnology , Chinese Academy of Tropical Agriculture Sciences, Haikou 571101, China; 3.Key Laboratory of Agriculture Biotechnology of Shihezi University, College of Life Sciences, Shihezi University,Shihezi 832003, China;4.College of Life Sciences, Beijing Normal University, Beijing 100875, China)

Abstract：DELLA protein is the key negative regulator of GA signal response. In order to study the effect of GA on development of cotton fiber, we cloned GhGAI gene, a member of DELLA gene family from the upland cotton EST database by electronic cloning method based on EST. According to the electronic cloning sequence, an integrate coding region was amplified from cotton( Gossypium hirsutum ) ovule cDNA. A protein band with molecular weight (Mr) of 62000 was obtained from the prokaryotic expression products by SDS-PAGE. The protein was then purified by His-tag affinity chromatography and used for multiple-cloned antibody preparation by rabbit immunization. Indirect ELISA test and the Western blot analysis showed that the anti-serum was in high immunocompetence and high specificity.
Key words：Gossypium hirsutum; GhGAI gene; polyclonal antibody; Western blot   [Full Text, 1952KB]