茶树基因芯片的研制和初步应用DevelopmentandPreliminary.PDF

2006 26 3166~170 Journal of Tea Science 1 2 1* 1 1 (1. / 310008 2. 310029) EST 1 680 cDNA 6 912 6 720 160 32 4×4 1 037 /cm2 3 2 PCR cDNA cDNA PCR S571.1; Q523 A 1000-369X200603-166-05 Development and Preliminary Application of cDNA Microarray of Tea Plant (Camellia sinensis) 1 2 1 1 1 ZHAO Li-ping , GAO Qi-kang , CHEN Liang *, WANG Xin-chao , YAO Ming-zhe (1 Lab for Tea Germplasm, Genetic Improvement and Molecular Biology, Tea Research Institute Chinese Academy of Agricultural Sciences; Key Laboratory of Tea Chemical Engineering, the Ministry of Agriculture, Hangzhou 310008, China; 2 Biological Macromolecular Research Laboratory of Analyzing and Measurement Center, Zhejiang University, Hangzhou 310029, China) Abstract: Totally, 1 680 genes obtained in our EST project were selected from the cDNA library of clone Longjing 43 to develop the first cDNA microarray of tea plant. Each gene in the microarray was duplicated. The cDNA microarray contains 6 912 dots, including 6 720 EST, 160 positive controls and 32 negative controls. One microarray was spotted two regions with 4×4 matrixes and it could be hybridized twice. The density of the microarray was 1037 dots per cm2 . Three tea clones with different contents of tea polyphenol were selected to conduct two sets of hybridization experiments with the microarray and the differently expressed ESTs were found. Then, two genes that were closely related to tea aroma were selected for real-time PCR analysis to validate the microarray data and validated the reliability of the cDNA microarray. The newly developed cDNA microarray could be applied in various tea research fields to mak