Chapter title |
Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants, Second Edition
|
---|---|
Chapter number | 22 |
Book title |
Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants, Second Edition
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3332-7_22 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3330-3, 978-1-4939-3332-7
|
Authors |
Singh, Rupesh Kumar, Hou, Weina, Franklin, Gregory, Rupesh Kumar Singh, Weina Hou, Gregory Franklin |
Abstract |
Hypericin, an important determinant of the pharmacological properties of the genus Hypericum, is considered as a major molecule for drug development. However, biosynthesis and accumulation of hypericin is not well understood. Identification of genes differentially expressed in tissues with and without hypericin accumulation is a useful strategy to elucidate the mechanisms underlying the development of the dark glands and hypericin biosynthesis. Suppression Subtractive Hybridization (SSH) is a unique method for PCR-based amplification of specific cDNA fragments that differ between a control (driver) and experimental (tester) transcriptome. This technique relies on the removal of dsDNA formed by hybridization between a control and test sample, thus eliminating cDNAs of similar abundance, and retaining differentially expressed or variable in sequence cDNAs. In our laboratory we applied this method to identify the genes involved in the development of dark glands and accumulation of hypericin in Hypericum perforatum. Here we describe the complete procedure for the construction of hypericin gland-specific subtracted cDNA library. |
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