Cloning of homeobox cDNA from the platyfish(Xiphophorus spp.)

• Main Authors: Hue-zu Li, 李蕙如
• Other Authors: Ku-yu Lu
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/66847195080950126805

碩士 === 東吳大學 === 微生物學系 === 93 === Homeobox genes were understood as a major body plan regulators. In Gordon-Kösswig Xiphophorus hybrid system, the pattern of black spots on the body of the purebred fish and hybrid offsprings always show the same body-site-specific expression characteristics. In order to understand the relationship between homeobox genes and Xiphophorus body-site-specific expression, in this study, the expression of different homeobox paralogs on different fish compartments were analysed, and a specific homeobox fragment was cloned and sequenced. The fish compartments under investigation include：dorsal fin, tail fin, skin, heart, liver, spleen, testicle and brain. Different pairs of primers were used in the RT-PCR reaction to analyse the homeobox gene expression. Finally, by using 3’-RACE (rapid amplification of cDNA ends), a clone named as 3R5 were cloned from mRNA preparation from spleen. 3R5 have nucleotide sequences indicated that the complement cDNA contained 912 nucleotide sequences. Nucleotide sequence analysis show that 3R5 fragment contains a distinct homeobox sequence and show high homology with the PDX-gene from Oryzias latipes（accession number ABO2 6976）and Danio rerio（accession number AFO3 6325）with 91 % and 84 % identity at nucleotide sequences level respectively, and 84 % identity at amino acid sequence level.