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Construction of Flax Male Sterile Gene Expression Vector

Author: LvQiuShi
Tutor: ZhangHui；SiQinBaTeEr
School: Inner Mongolia Agricultural University
Course: Crop Genetics and Breeding
Keywords: Flax male sterility A9 promoter EPSPS gene Sequence analysis Vector construction
CLC: S563.2
Type: Master's thesis
Year: 2011
Downloads: 19
Quote: 0
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Abstract


In this study, a dominant nuclear male infertility flax material , difficult to use for the study material existence , based on preliminary studies , the use of biotechnology , build two flax male sterility expression vector for the creation of male sterility in flax new material basis. The specific content and the results are as follows : (1) in accordance with the relevant literature , design synthesis in Arabidopsis tapetum - specific expression of the A9 gene promoter -specific primers and Arabidopsis genomic DNA as a template for PCR amplification . Sequence determination and analysis revealed that the Arabidopsis thaliana A9 promoter successfully isolated . ( 2 ) According to the literature , the design and synthesis of the EPSPS gene -specific primers and transgenic soybean genomic DNA as a template for PCR amplification . Sequencing and analysis results show that the EPSPS gene successfully isolated . (3) using the digestion and connection technology, the promoter with the CaMV35S pHANNIBAL-172 and pHANNIBAL-944 A9 promoter replacement, constructed pHANNIBAL-A9-172 and pHANNIBAL-A9-944 vector . NPT Ⅱ gene in pART27 replace EPSPS gene was successfully constructed pART27-EPSPS carrier . (4) by Not I cleavage point , the two \carrier pART27-EPSPS construct MS2-F and herbicide-resistant gene 2 flax male sterility gene chain flax male sterility gene expression vector . The import of the carrier flax , MS2-F gene fragment in reverse order , under the guidance of the tapetum -specific A9 promoter expression into a double-stranded RNA inhibition of endogenous gene expression of MS2-F , resulting flax pollen sterility . Herbicide resistance gene as a selectable marker can be used spraying glyphosate methods screening flax male sterility in transgenic plants .

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CLC: > Agricultural Sciences > Crop > Economic crops > Fiber crops > Fiber crops > Flax
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