Cloning and Analysis of Multidrug Resistance-associated Protein Homolog in White Botton Mushroom

• Main Authors: Hsueh-Tze Shih, 施雪姿
• Other Authors: Yueh-Long Chang
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/05216548886234441025

碩士 === 國立嘉義大學 === 農業生物技術研究所 === 94 === Abstract In plant and animal cells, they are able to separate and eliminate toxicants from the cytoplast. In an instance of some lipiotoxicants, they can conjugate with glutathione to form GS-X or GSSG; and then these compounds could be eliminated by GS-X pump of MgATP-dependent transporter. In Saccharomyces cerevisiae, yeast cadmium factor（YCF1） belongs to the family of GS-X pump of MgATP-dependent transporters. More studies have shown that it is a number in the ATP binding cassette（ABC）transporter superfamily and belongs to the subfamily of Multidrug resistance-associated protein（MRP）. YCF1 is capable of binding with heavy metals and drugs which subsequently transported into vacuole; and it is 43％ identical （63％ similar）at the amino acid level with MRP1 in human. The recent years, edible mushroom has become more popular in fresh and processed usages. Some reports showed that the fruiting body of some mushrooms contains a high amount of heavy metals; therefore, in the mushroom genome, it could possess genes for the absorption and resistance of heavy metals. For cloning the YCF1-similar gene in Agaricus bisporus, the degenerate primes were designed from the conserved domains of YCF1 gene. In result, a fragment of YCF1-homologous gene was successfully amplified and identified by sequencing. In addition, a cDNA library of white button mushroom was developed. It will be used for the identification of full length of YCF1-similar homologs and its gene family. Keywords：Heavy metal、Agaricus bisporus、Saccharomyces cerevisiae cadmium factor（YCF1）、Multidrug resistance-associated protein（MRP）subfamily、Degenerate primer、cDNA library。