Yunnan red pear PybHLH gene as well as prokaryotic expression vector and application thereof

A prokaryotic expression and gene technology, applied in the field of genetic engineering, can solve problems such as less research, and achieve the effect of strong advantages, accurate experimental results, and convenience for subsequent experiments.
CN103146709AInactive Publication Date: 2013-06-12KUNMING UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
KUNMING UNIV OF SCI & TECH
Publication Date
2013-06-12
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a Yunnan red pear PybHLH gene through anthocyanin biosynthesis, trichome development and a salt stress regulatory protein and a prokaryotic expression vector thereof. The PybHLH gene is cloned from red fruit peel of Yunnan red pear NO.1 by using a RT-PCR (reverse transcription-polymerase chain reaction) technology, then the prokaryotic expression vector pGEX-4T-PybHLH is constructed, PybHLH protein is expressed in escherichia coli Rosetta (DE3) by pGEX-4T-PybHLH vector and then is purified, and Yunnan red pear PybHLH purified protein is obtained; the prokaryotic expression vector of the PybHLH gene is applied to preparing a pybHLH specific antibody, and the obtained pybHLH specific antibody is also used for researching the interaction of the protein and protein as well as the protein and DNA(deoxyribonucleic acid) and the detection to PybHLH transgenic plant. The obtained pybHLH specific antibody contains glutathione S transferase (GST) and is applied to researching the interaction of the protein through Far Western blotting and co-immunoprecipitation and accurately separating and obtaining the protein and DNA segment which are interacted with PybHLH.
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Description

technical field

[0001] The invention belongs to the field of genetic engineering, and specifically relates to a Yunnan red pear anthocyanin biosynthesis, trichome development and salt stress regulation protein gene PyBH The prokaryotic expression thereof and the application of the prokaryotic expression vector in the preparation of PybHLH protein and specific antibody. Background technique

[0002] Red peel is an important trait index in pear molecular breeding. Due to its unique geographical and climatic conditions, Yunnan Province has more germplasm resources of red skin pears. Since 1986, four cultivars, Zaobaimi, Meirensu, Mantianhong and Yunhongli No. 1, have been bred successively. However, except for Yunhongli No. 1, the other three varieties will have poor coloration or even no coloration due to climate change. Therefore, it is necessary to study the coloration mechanism of red pear peel. Previous studies have shown that the synthesis of plant anthocyanins is regul...

Claims

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