毛竹微型颠倒重复序列转座子PhTourist1的克隆与分析

doi:10.11707/j.1001-7488.20150515

Abstract

Abstract:

【Objective】 Miniature inverted-repeat transposable elements (MITEs), some of which are known as active non-autonomous DNA transposons, can jump by transposases encoded by the autonomous DNA transposons. It has been reported that the transposition of MITE influences the expression of host's genes. In order to study the structural characteristics and transposition characteristics of MITE, and to analyze the impact of the transposition of MITE on expression of its near genes in Phyllostachys edulis genome, the Tourist-like MITE PhTourist 1 which was homologous with mPing from rice was chosen to analyze the role of transposons in genome polymorphism formation and gene expression regulation in P. edulis. 【Method】TIR (terminal inverted repeat) and TSD (target site duplication) of PhTourist 1 were identified and the insert preference of PhTourist 1 was investigated using software of MEGA5.1, DNAStar and WebLogo 3-Create. Then the insertion time and distribution of PhTourist 1 in the P. edulis genome were calculated. PhTourist 1 insertion sites in the P. edulis genome were tested by PCR in the 24 seedlings to test their insert polymorphisms. With the real-time fluorescence quantitative PCR (RTFQ PCR) technique, the expression level of gene downstream PhTourist 1-3 was analyzed with PhTourist 1-3 exists or not.【Result】 30 copies of PhTourist 1 were discovered. Their TIR(GGCCAGTCTCAATG),TSD(TWA) and insert preference sequence((C/T)T(C/A)T(T/A)A(G/T)A(A/C)) identified with bioinformatics tools were consistent with previous reports. The insert time of PhTourist 1 in P. edulis genome was relatively concentrated. PhTourist 1 also preferentially inserts near genes. Each of the insertion sites of PhTourist 1 was verified correct, while only PhTourist 1-3 has insert polymorphism among the 30 copies. There was no footprint at the excision site of PhTourist 1-3 which was in cosistence with mPing. The analysis by RTFQ PCR of the expression level of a downstream gene PH01000402G0860 of PhTourist 1-3 revealed an increase by 8.04 times with the missing of PhTourist 1-3 than the existing of PhTourist 1-3. 【Conclusion】 PhTourist 1-3, one of the identified PhTourist 1 homology with mPing in the study, could jump in the genome of P. edulis. It was the first active Tourist-like MITE discovered in P. edulis and had significant influence on expression of its downstream genes. The results showed that transposons as an important part of P. edulis genome could participate in the formation of gene polymorphism, regulate the gene expression and involve in the regulatory of host growth and development.

Key words: Phyllostachys edulis, Tourist-MITE, mPing, transposition, gene expression

CLC Number:

S718.46

Hu Hui, Zhou Mingbing, Yang Ping, Tang Dingqin. Cloning and Analysis of Miniature Inverted Repeat Transposable Elements PhTourist 1 from Phyllostachys edulis[J]. Scientia Silvae Sinicae, 2015, 51(5): 127-134.

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Cloning and Analysis of Miniature Inverted Repeat Transposable Elements PhTourist 1 from Phyllostachys edulis

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