Cloning and Functional Analysis of the Genes that Enhance Bioluminescence of Photobacterium leiognathi lux Operon

• Main Authors: FangChun Sun, 孫芳君
• Other Authors: JueyWen Lin
• Format: Others
• Language: zh-TW
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/86751091698730687606

碩士 === 國立中興大學 === 分子生物學研究所 === 87 === Plasmid pL741 carries a ~15 kb HindIII partial digested Photobacterium leiog-nathi genomic DNA in pACYC 184. However, the P. leiognathi lux operon cloned in pL741 shows dim bioluminescence in E. coli; it suggested that the lux operon required regulatory gene(s) to induce or enhance the gene expression. In trans complementa-tion bioluminoassays in vivo were used to construct P. leiognathi genome library for cloning the specific genes, which enable to enhance bioluminescence of the lux operon. pXY5 and pXY6 were isolated from two of the selected bright clones. Nucleotide sequences of the P. leiognathi genes carried on pXY5 and pXY6 were determined, and the genes were identified. The gene orders of the genes are -ufoI-ufoII-WTRR&R-gltS-WTI-WTIIR and torA-R&R-torCR on pXY5 and pXY6, respectively (R&R: regulatory region; W: transcription terminatior). Apparently ufoII, torA, torC, gltS genes are not the specific regulatory genes of the lux operon; but functional analyses show that ufoII, torA, torC genes enable to enhance biolumine-scence. It elicits that not only the specific regulatory genes enable to enhance bioluminescence of the lux operon, also many other genes enable to do. However, ufoII, torA, torC genes can enhance bioluminescence of the lux operon in E. coli, but the enhance mechanisms are not clear yet.