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Construction of Large DNA Fragment Cloning Vectors and Rice Transformation Based on Transformation-competent Artificial Chromosome (TAC) Vector

Author: LinMeiKun
Tutor: WangFeng
School: Fujian Agriculture and Forestry University
Course: Biochemistry and Molecular Biology
Keywords: Rice Polygenic Transformation artificial chromosome Genetic transformation
CLC: S511
Type: Master's thesis
Year: 2009
Downloads: 130
Quote: 1
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Abstract


In this study, the plant can be transformed artificial chromosome (TAC) vector technology used in the rice four kinds transporter protein gene sucrose transporter protein gene (ossut5z), cell wall invertase gene (oscin3), monosaccharide transporter gene (osmst4) and ATP / ADP transporter protein gene (oaat) is multi - gene vectors and genetic transformation . The vector was constructed through the backbone vector to pYLTAC to build four genes related to the control assimilation product transportation such as OsSUT5Z, OsMST4, OsCIN3 OAAT same expression vector , were constructed the carrier pTAC747H - CIN3 - MST4 , pTAC747H the - OAAT - SUT5 and pTAC747H - OAAT - SUT5 - CIN3 - MST4 total insert size is about 30kb . Laboratory original transformation system based on preliminary exploration through Agrobacterium -mediated transformation of the large fragment of multi - gene vector conversion efficiency factors , including different rice genotypes , different plant medium , different Agrobacterium infection callus organized way , and other factors , so as to effectively improve the efficiency of the conversion of large fragment . Three expression vectors into Agrobacterium mediated transformation Minghui 86 and Nipponbare callus of mature embryos , which turn Minghui 86 conversion rate were 12.5% ??, 11.3% and 8.4 % , respectively ; transfer Nipponbare The conversion rate of 11.7% , 9.8% and 8.7% , respectively . 8,5 and 4 cloned transgenic rice plants , molecular detection showed that these cloned exogenous genes had been integrated into the rice genome .

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CLC: > Agricultural Sciences > Crop > Cereal crops > Rice
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