Cloning, Identification And Analysis Characteristics Of LINEs And Ty3-gypsy Retrotransposons From Bamboo

Retrotransposons were widely distributed in plant, which of copy and paste mechanism was veryimportant to the evolution of plant genomic. Combine molecular technology with bioinformatic, ourresearch analyzed the structure and characteristic of LINEs and LTR, obeserved the active of LTR inArabidopsis thaliana.1.45sequences were obtained from Phyllostachys heterocycla cv.pubescens, Sasafortunei(VanHoutte)Fiori, Phyllostachys heterocycla（Carr.） Mit ford, respectively, which of lengh varied from407-446bp. Nucleotide sequnces comparasion showed a high level of divergence, whereas amino acid comparasion demostrated that DomainII、DomainIII and DomainIV were conservative.Based on the amino acid sequnce, the phylogenetic tree indicated that sf-2and pp-29are in th-e same group, which suggested that horizontal transmission may occur in LINEs.2. A LTR Retro-transposon named Ph-LTR1was cloned from Phyllostachys heterocyclacv.pubescens. Sequnecse analysis showed that the full length of Ph-LTR1was5350bp, which coding apolypeptide of1534amino acids and contain gag gene、RT gene、RH gene plus int gene respectively.Ph-LTR1belonged to Ty3-gypsy Retro-transposons which was based on the analysis of the genesequence. Structure analysis and comparasion with other elements from Phyllostachys heterocyclacv.pubescens showed that gene fragment deficience exists in most elements, which suggested that thePhyllostachys heterocycla cv.pubescens gemomes would prefer to keep the relatively conservativedomain over evolutionary time. Ph-LTR1was a possibly active retrotransposon judged from theevaluation of insertion time.3. Through ligating Ph-LTR1to the multiple clone sites of pCAMBIA1301, the plant expressionvectors pC1301-LTR1was constructed and transformed to Arabidopsis thaliana with anAgrobacterium-mediated floral-dip method. Trans-formants were identified and detected of theexpression of the transposon in Arabidopsis thaliana, and this study found that transcription has beenhappened in gag gene、RT gene and int gene, respectively. Those obtained transgenic lines would be ofgreat help to the study, which would lay a basis for the future research about unique escision and geneticstability in Arabidopsis thaliana genome.