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Vitis CBF cold response pathway gene cloning and expression vector plants

Author: LiXiaoZuo
Tutor: LiHaiYan
School: Jilin Agricultural University
Course: Crop biotechnology
Keywords: Wild grape (Vitis amurensis) Low temperature reaction pathway CBF genes Clone Construction of plant expression vectors
CLC: S663.1
Type: Master's thesis
Year: 2008
Downloads: 176
Quote: 0
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Abstract


Arabidopsis CBF (CRT / DRE binding factor) transcription factor in a cold response pathway plays an important role , a CBF transcription factor can activate a number of downstream genes associated with resilience expression . Although there are multiple plants cold response pathway , but for the most in-depth study CBF pathway , the most precise , so CBF gene family is the preferred plant cold molecular breeding objectives . In this paper, PCR from the wild grape (Vitis amurensis) was isolated and cloned two new CBF gene ( gene bank registration number DQ517297 and DQ517298), were named VaCBF2 and VaCBF3; also constructed two gene plant expression vector . The main results are as follows : 1. Published in GenBank by some plants to compare CBF homologous genes conserved region in its design one pair of degenerate primers , using PCR from cDNA Vitis get two sizes were 536bp and 527bp of middle CBF gene fragments were cloned and sequenced . 2 In the two intermediate fragment area were designed two pairs of specific primers , using inverse PCR method Vitis CBF gene 5 'and 3' noncoding region was cloned , the results were obtained two lengths of about 1200bp and 1000bp fragment . Intermediate fragment length predicted genes spliced ??region . 3 in the region predicted full-length non-coding region were designed a pair of specific primers for amplification of Vitis genome were obtained 954bp and 885bp fragments . Confirmed by sequencing the resulting sequence is the length of CBF genes were named as VaCBF2 and VaCBF3. 4. Application software will VaCBF2 and VaCBF3 DNAMAN length with Genebank published several plants CBF gene amino acid homology was found VaCBF2 and VaCBF3 all have conserved and CBF family of AP2 series of features , which shows that we have successfully cloned VaCBF2 and VaCBF3. 5. use of semi- quantitative RT-PCR method VaCBF2 and VaCBF3 expression analysis . 6 Use the Gateway technology to build VaCBF2 and VaCBF3 plant expression vector into the two expression vectors in Agrobacterium EHA0105 .

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CLC: > Agricultural Sciences > Gardening > Fruit trees gardening > Berries > Grapes
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