03103nas|a2200409 i 4500 20260817172937.0 020215s2001 th uzm rtt 00| a1eng d .b1184596x AIT Thesis no. BP-01-05 Nguyen Hong Nhung Cloning of fungal chitin deacetylase gene into bacterial Escherichia coli Bangkok : Asian Institute of Technology, 2001 40 leaves Thesis ; no. BP-01-05 A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science, School of Environment, Resources and Development Thesis (M.Sc.) - Asian Institute of Technology,2001 Chitin deacetylase(CDA)gene from Colletotrichum lindemuthianum UPS9 has been used in this study. The gene cloned into shuttle vector pPIC9K has been used as a source of CDA gene throughout the study. Large quantity of gene was obtained allowing its multiplication in component E.coli TOPlOF'. QIAGEN plasmid midi Kit was used for the extraction and purification of the DNA. The gene was integrated into a another shuttle vector pPIC3.5K in its multiple cloning site. The plasmid vector pPIC3.5Kb is 9.0 kb long plasmid earring ampicillin and kanamycin resistant markers. It can be multiplied in E.coli as well as in Picia pastoris. In E.coli, it remains extrachromosomally and multiplies independently. It remains as a low copy number plasmid. On the other hand , in Pichia pastoris, it intergrates into genome via homologous recombination and results in stable transfmmation. In this research work, CDA gene has been intergrated in between EcoRI and Natl sites of multiple cloning sites. Recombinant plasmid was transformed into competent E.coli TOPIOF' intermediate host by calcium chloride method in order to obtain the plasmid in large quantity. The integration of CDA gene was confimed by restriction digestion analysis followed by gel electrophoretic migration and size comparison. Chitin Escherichia coli Stevens, Willem Frans, Chairperson Suwalee Chandrkrachang, Examination committee Sunee Nitisinprasert, Examination committee Macaranas, Julie M., Examination committee Swedish International Development Cooperation Agency (Sida), Scholarship donor Asian Institute of Technology. Thesis ; no. BP-01-05 Full-Text http://203.159.5.9/ait-thesis/detail.php?q=B07827 .b1184596x mnait u 240329 0 020215 m a u 0 mnait mnarc 22 40 Barcode : 30050120683056 CREATED : 2013-06-09 RECORD # : i12740019 LPATRON : 0 LCHKIN : - # RENEWALS : 0 # OVERDUE : 0 IUSE3 : 0 TOT CHKOUT : 0 TOT RENEW : 0 Barcode : 30050120350888 CREATED : 2016-10-06 RECORD # : i13029605 LPATRON : 0 LCHKIN : - # RENEWALS : 0 # OVERDUE : 0 IUSE3 : 0 TOT CHKOUT : 0 TOT RENEW : 0 18113 18113 0 0 0 0 MAIN MAIN mnait 2026-08-17 50.00 0 AIT Thesis no. BP-01-05 30050120683056 2026-08-17 17:29:37 3 2026-08-17 22 0 0 0 0 MAIN MAIN mnarc 2026-08-17 0 AIT Thesis no. BP-01-05 30050120350888 2026-08-17 17:29:37 2026-08-17 40