03206nas|a2200445 i 4500 20260817172937.0 020717s2000 th u m rtt 00| a1eng d .b11861435 AIT Thesis no. BP-00-10 Shrestha, Binesh Cloning of fungal chitin deacetylase gene into yeast Pichia pastoris Bangkok : Asian Institute of Technology, 2000 82, [2] leaves Thesis ; no. BP-00-10 A thesis submitted in partical fulfillment of the requirements for the degree of Master of Science, School of Environment, Resources and Development Thesis (M.Sc.) - Asian Institute of Technology, 2000 Chitin deacetylase (CDA) gene has been amplified from Colletotrichum lindemuthianum UPS9 DNA and cloned into one of the E.coli-Pichia pastoris shuttle vector (pPlC9K). The sequence analysis of cloned gene showed 98% homology with that of C. /indemuthianum ATCC56676. The gene was cloned into pPIC9K in frame with the asecretion signal factor present in the vector. Furthermore, the gene was transformed into Pichia pastoris expression host in order to obtain overexpressing strains. Two strains of Pichia namely GS115 (fast methanol utilizing, Mut+) and KM71 (slow methanol utilizing, Mut5 ) were selected as a host for the exrpession of CDA gene. Pichia transformants were selected on the basis of Histidine synthesizing phenotype and were screened for transformants carrying multi copy inserts of CDA gene on the basis of degree of resistance to G418 (Geneticine ). Transformants carrying resistance upto 3 mg/ml of G418 were screened out which might carry several copies of cloned CDA gene. The expression of cloned CDA gene in Pichia transformants were studied by analysing culture supernatant on SOS-PAGE for excreted protein expression. Molecular cloning Chitin Stevens, Willem Frans, Chairperson Hegarat, Francoise Le, Co-Chairperson Suwalee Chandrkrachang, Examination committee Pakorn Nuchnoi, Examiantion Committee Sunee Nitisinprasert, Examination committee H.M. the King of Thailand, Scholarship donor Asian Institute of Technology. Thesis ; no. BP-00-10 Full-Text http://203.159.5.9/ait-thesis/detail.php?q=B06980 .b11861435 mnait u 240402 0 020717 m a u 0 mnait mnait mnarc 20 40 Barcode : 30050120682876 CREATED : 2001-02-17 RECORD # : i12291018 LPATRON : 1015270 LCHKIN : 2003-02-21 # RENEWALS : 0 # OVERDUE : 0 IUSE3 : 0 TOT CHKOUT : 4 TOT RENEW : 0 Barcode : 30050120682850 CREATED : 2001-02-17 RECORD # : i1229102x LPATRON : 1016802 LCHKIN : 2004-02-29 # RENEWALS : 0 # OVERDUE : 0 IUSE3 : 0 TOT CHKOUT : 5 TOT RENEW : 10 Barcode : 30050120350821 CREATED : 2016-10-06 RECORD # : i1302968x LPATRON : 0 LCHKIN : - # RENEWALS : 0 # OVERDUE : 0 IUSE3 : 0 TOT CHKOUT : 0 TOT RENEW : 0 18114 18114 0 0 0 0 MAIN MAIN mnait 2026-08-17 50.00 0 AIT Thesis no. BP-00-10 30050120682876 2026-08-17 17:29:37 1 2026-08-17 20 0 0 0 0 MAIN MAIN mnait 2026-08-17 50.00 0 AIT Thesis no. BP-00-10 30050120682850 2026-08-17 17:29:37 2 2026-08-17 20 0 0 0 0 MAIN MAIN mnarc 2026-08-17 0 AIT Thesis no. BP-00-10 30050120350821 2026-08-17 17:29:37 2026-08-17 40