Plasmid curing experiments showed that the Caprylate-Esterase (CE) gene of Lactobacillus casei subsp. casei (LLG) is located on the LLG chromosome rather than on a plasmid. The enzyme was purified and shown to be a monomer with an apparent Mr of 32,000 on sodium dodecyl sulfate-polyacrylamide gels. The N-terminal end of the CE protein was microsequenced. A synthesized 29-mer oligonucleotide deduced from L. casei codon usage and the N-terminal peptide sequence was used as probe against an LLG genomic library. Library screening yielded one positive clone (pLLG1435) with an insert of 8 kb. Southern analysis of pLLG1435 showed that the oligonucleotide probe strongly hybridized with the 1.9 kb PstI/SphI fragment found within its insert. Th fragment was subcloned and its DNA sequence was determined and found to be closely related to the phosphoribosylformylglycinamidine synthase II gene of Bacillus subtilis. The gene was completely sequenced.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.61201 |
| Date | January 1991 |
| Creators | Gu, Zhengming |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Microbiology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001275460, proquestno: AAIMM74884, Theses scanned by UMI/ProQuest. |
Page generated in 0.0022 seconds