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Expression Analysis of the Promoter of ANK and Pita in Transgenic Rice

Author: LiXiuJuan
Tutor: HeShuiLin
School: Fujian Agriculture and Forestry University
Course: Biochemistry and Molecular Biology
Keywords: Promoter Ankyrin Pita Quantitative analysis of GUS Rice
CLC: Q943.2
Type: Master's thesis
Year: 2011
Downloads: 50
Quote: 0
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Abstract


Gene expression in large part by its promoters and its specific cis-acting elements of the interaction of the decision to analyze the law regulating the expression of the promoter gene function is an important way to clear. Plants in the long process of evolution, the development of inducible resistance, stress resistant pathogenic bacteria will be restricted to certain conditions in order to reduce unnecessary waste of materials and energy and reduce energy sterilization of toxic secondary metabolites of plants cells themselves toxic effects. Therefore, the analysis of plant disease resistance gene promoter not only help elucidate the function of plant disease resistance genes, but also for plant genetic engineering to improve the expression of exogenous resistance genes provide application value inducible promoter. In view of this, the project by obtaining promoter GUS reporter gene vector T1 generation of transgenic rice lines analyzed Pita rice blast resistance genes and pathogenic bacteria showed significant response Stress the role of rice ankyrin gene promoter , aims to analyze the gene expression pattern in rice, clear as inducible promoter in rice disease resistance genes possibilities. The main results are as follows: 1, rice ankyrin gene promoter Functional Analysis: According to the online prediction of rice primers designed ankyrin gene from rice (japonica variety Nipponbare) to amplify the gene promoter fragment, called OsANKp, will its promoter sequence in the promoter cis-acting elements forecast website PALCE for analysis, predicted that more than 10 possible components. Constructed OsANKp :: GUS expression vectors, the use of Agrobacterium-mediated genetic transformation of rice was transformed to obtain a certain number of transgenic lines. Transgenic plants may be subject to mechanical damage, leaf roller, rice blast treatment induced expression and growth over time, GUS expression intensity also increases. 2, rice blast resistance gene Pita promoter Functional Analysis: According to the online prediction of rice Pita gene primers designed from rice (japonica variety Nipponbare) to amplify the gene promoter fragment, called OsPITAp, start it sequences on the promoter cis-acting elements forecast website PALCE on the analysis, forecast to more than 20 possible components. Constructed OsPITAp :: GUS expression vectors, the use of Agrobacterium-mediated genetic transformation of rice was transformed to obtain a certain number of transgenic lines. OsPITAp the T1 generation transgenic plants, the stems and leaves of GUS gene expression is not found, the root of the expression of a small amount, hulled seeds and chaff are also a small number of GUS gene expression in transgenic plants of this study to mechanical damage, rice blast no acknowledge; GUS quantitative analysis MeJA, NaCl treatment can significantly increase the OsPITAp :: GUS expression; synthetic small molecule compounds elicitor INA (2,6 - dichloro-isonicotinic acid), and n-[(2s, 3r) -3-amino-2-hydrcxy-4-phenylbutyryl]-L-leucin (phenylbutyrate inhibin, Ubenimex) shoots after treatment can enhance the expression of GUS.

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CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
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