An Annealing Control Primer (ACP) System Used for the Isolation and Identification of Copper-Induced Genes in Alfalfa Leaves
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- 2016.11.02
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서지정보
ㆍ발행기관 : 한국초지조사료학회
ㆍ수록지정보 : 한국초지조사료학회지 / 36권 / 3호
ㆍ저자명 : Ki-Won Lee, Sang-Hoon Lee, Ki-Yong Kim, Hee Chung Ji, Hyung Soo Park, Tae Young Hwang, Gi Jun Choi, Md Atikur Rahman
목차
Ⅰ. INTRODUCTION
Ⅱ. MATERIALS AND METHODS
1. Plant material and copper treatment
2. Isolation of RNA, first strand cDNA synthesis andACP-based GeneFishing
3. Gene cloning sequencing and bioinformatics analysis
Ⅲ. RESULTS AND DISCUSSION
Ⅳ. CONCLUSION
Ⅴ. ACKNOWLEDGEMENTS
Ⅵ. REFERENCES
영어 초록
Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.
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