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红巴梨f3h基因的克隆及植物表达载体的构建(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2005年02期
Page:
65-68
Column:
研究论文
publishdate:
2005-02-20

Article Info:/Info

Title:
Cloning f3h Gene from "Red Ballet" Fruit and Its Construction of Plant Express Vector
Article ID:
1000-2006(2005)02-0065-04
Author(s):
ZHANG Da-sheng1 CUI Li-jie2 WANG Jing-ming1 WU Shao-hua2*
1. School of Life Science, Xuzhou Normal University, Xuzhou 221116, China; 2. College of Horticulture, Fujian Agricultural & Forestry University, Fuzhou 350002, China
Keywords:
Anthocyanin f3h gene Clone Vector
Classification number :
Q813
DOI:
10.3969/j.jssn.1000-2006.2005.02.016
Document Code:
A
Abstract:
The fragment of f3h gene of anthocyanin biosynthesis was amplified by RT-PCR method using total RNA from ripping "Red Ballet" fruit peel as template and oligonucleotides designed from the published sequence of kernel fruits as primers. Sequence analysis showed that this fragment is full-length cDNA of f3h gene and it contained 1 095 base pairs. Sequence comparison revealed that the f3h gene of "Red Ballet" shared homologies of 92%, 83% and 82% with those of "Fuji" apple, Petunia and Arabidopsis isolates., respectively. Then the f3h gene was digested by EcoR V and Bgl Ⅱ endoenzymes and 1 095 bp fragment was recovered. Vector pBI121 and 1 095 bp fragment,were connected using ligase. Reconstituted plasmid pBI12l included the anticipated fragment of f3h gene was obtained and was proved by antibiotic selection and PCR analysis. The reconstituted plasmid is using for the further study.

References

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Last Update: 2013-05-20