Gene Cloning of N-acetyltransferase and Biodegradation of 3,4-dichloroaniline of Trichoderma virens FT-333

• Main Author: 蔡建韋
• Other Authors: Ruey-Shyang Chen
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/06490084960657462247

碩士 === 國立嘉義大學 === 生化科技學系研究所 === 105 === Trichoderma spp. are widely used in agriculture, with its high adaptability and tolerance in soil, and the abilities to promote growth and resistance of plants. Previous studies have shown that Trichoderma spp. can degrade 3,4-dichloroaniline (3,4-DCA), a toxic compound from pesticide derivatives in soil, by N-acetyltransferase (NAT). In this study, the specific primers were used to amplify the NAT gene of Trichoderma FT-333. The results of sequence alignment showed that the identity of NAT1 and NAT2 between FT-333 and gene sequences (BN001413.1 and BN001414.1) in the GenBank were 93% and 99%, respectively. Inverse PCR (IPCR) was further performed to clone the full sequence of NAT gene. HPLC was also used to investigate the degradation of 3,4-DCA into acetyl 3,4-DCA by NAT in the medium innoculated FT-333. More, total protein from FT-333 culture were purified by the ammonium sulfate precipitation, and the protein ranged from 26 to 35 kDa were collected. These proteins were found to degrade 3,4-DCA into acetyl 3,4-DCA, suggested that the molecular weight of NAT of FT-333 was between 26-34 kDa.