Totally, 1β680 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 6β912 dots, including 6β720 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 make the high-throughput detection in the gene expression profiling.
ZHAO Li-ping
,
GAO Qi-kang
,
CHEN Liang
,
WANG Xin-chao
,
YAO Ming-zhe
. Development and Preliminary Application of cDNA Microarray of Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2006
, 26(3)
: 166
-170
.
DOI: 10.13305/j.cnki.jts.2006.03.002
[1] Schena M, Shalon D, Davis RW, et al. Quantitative monitoring of gene expression patterns with a complementary DNA microarray[J]. Science, 1995, 270(20): 467~470.
[2] Wang RC, Gueler K, Labrie ST et al. Genomic analysis of a nutrient response in Arabidopsis reveals diverse expression patterns and novel metabolic and potential regulatory genes induced by nitrate[J]. The Plant Cell, 2000(12): 1491~1509.
[3] Narusaka Y, Narusaka M, Seki M, et al. The cDNA microarray analysis using an Arabidopsis pad3 mutant reveals the expression profiles and classification of genes induced by Alternaria brassicicola attack[J]. Plant Cell Physiology, 2003, 44(4): 377~387.
[4] 马智华,万立华,晏珩,等. 利用基因芯片技术研究电击伤后心肌组织基因表达谱的改变[J]. 第三军医大学学报, 2005, 27(15): 1561~1563.
[5] Daborn PJ, Yen JL, Bogwitz MR, et al. A single P450 allele associated with insecticide resistance in Drosophila[J]. Science, 2002, 297(27):2253~2256.
[6] 黄迎春, 孙春昀, 冯红, 等. 利用基因芯片检测转基因作物[J]. 遗传, 2003, 25(3): 307~310.
[7] 李瑶, 裘海燕. 用基因表达谱芯片研究人正常肝和肝细胞癌中差异表达的基因[J]. 遗传学报, 2000, 27(12): 1042~1048.
[8] 伍亚舟, 张彦琦, 黄明辉, 等. 基因芯片表达数据的标准化策略研究[J]. 第三军医大学学报, 2004, 26(7): 594~597.
[9] 赵丽萍, 陈亮, 王新超, 等. 茶树新梢不同叶片中β-葡萄糖苷酶和β-樱草糖苷酶基因表达的实时定量PCR分析[J]. 茶叶科学2006, 26(1): 11~16.
[10] Moore S, Vrebalov J, Payton P, et al. Use of genomics tools to isolate key ripening genes and analyse fruit maturation in tomato[J]. Journal of Experimental Botany, 2002, 53(377): 2023~2030.
[11] Kim H, Snesrud EC, Haas B, et al. Gene expression analyses of Arabidopsis chromosome 2 using a genomic DNA amplicon microarray[J]. Genome Research, 2003(13):327~340.
[12] Chen L, Zhao LP, Gao QK.Generation and analysis of expressed sequence tags from the tender shoots cDNA library of tea plant (Camellia sinensis)[J]. Plant Science, 2005, 168(2): 359~363.
浙公网安备 33019902000101号 
