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Cloning and Expression of Pinb Gene in Aegilops bicornis

CHEN Ming-jie~1,FANG Ti~2,LIU Yong~1,HUANG Wei-wei~1,YANG Guang-xiao~1,HE Guang-yuan~ 1* (1 China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory,Huazhong University of Science and Technology,Wuhan 430074,China;2 Biotechnology Base,College of Life Science,Wuhan University,Wuhan 430072,China)  
According to the reported conservative sequences of Pinb gene,a pair of specific primers were designed to amplify,clone and sequence Pinb gene of DNA genome in Aegilops bicornis (SS) and a new allele of Pinb was found.The allele was 360 bp long,encoded 119 amino acid residuals,corresponded with the mature protein structure region of PinB in cereal crops, had its specific tryptophan motif of WPTKWWK and contained five disulphide bonds formed by ten cysteines.In nucleitide and amino acid sequences,it is 93.1% and 91.6% as homologous as Pinb-D1a of T.aestivum cv.Capitole,respectively.It was proved by RT-PCR that Pinb gene expressed itself in grain endosperm.The results indicated that Aegilops bicornis contained the gene regulating grain hardness that greatly differed from that in wheat so that it could provide rich genetic resources for wheat quality improvement.
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