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The Cloning of the Cytosolic Glyceradehyde-3-phosphate Dehydrogenase(GADPH)Gene(TaGapCl)and Its Expression and Sequence Analysis in Iron-deficient Stress in Wheat

Author: LiZuo
Tutor: YinLiPing；QiuZeSheng
School: Capital Normal University
Course:
Keywords: Subtractive Hybridization in situ hybridizationof plaque Glyceradehyde-3-phosphate Dehydrogenase (GADPH) Iron-deficient stress Northern blotting Southern Blotting
CLC: S512.101
Type: Master's thesis
Year: 2000
Downloads: 119
Quote: 1
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Abstract


Fourteen clones were screened from the iron梔eficient cDNA library in wheat by hybridization using the specific probe obtained from the subtractive hybridization. Furthermore, these clones were identified by F. coil blue: white screening for plaque and PCR, then purified and recircularized into pMD18桾 vector etc. Five fragments of 0. 2?. 9kb in length were sequenced, one of which had elevated levels of transcription under iron梔eficient conditions. The sequence analysis revealed that it is a 812bp in length cDNA and the deduced amino acid sequence had significant identify with the cytosolic glyceradehyde?梡hosphate dehydrogenase (GADPH) gene (G~pc~ of other plants in the detabase, such as barley, maize, rice, Arabdopsis thaiiasa as well as tomato, tobacco and pea etc. So it is named as TaGapGl. Northern blot shows that this gene has the enhanced expression in iron梔eficient stress in the root of wheat and there has the similar result in the leaves. The result of Southern blot indicates that TaGapCl was a single copy belonged to the nucleus genome of wheat.

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