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Method for cloning promoter sequence of CcPIP in hickory

A technology of promoter sequence and hickory, applied in the field of bioengineering, can solve the problems of high test cost, high test technical requirements, unsatisfactory results, etc.

Inactive Publication Date: 2014-03-26
ZHEJIANG FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are the following defects: 1) DNA circularization is easily generated in the PCR process, resulting in non-specific amplification, resulting in poor amplification efficiency and often unsatisfactory results; 2) too long enzyme-cut fragments will also reduce the amplification efficiency, High test cost; 3) heavy workload and high experimental technical requirements

Method used

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  • Method for cloning promoter sequence of CcPIP in hickory
  • Method for cloning promoter sequence of CcPIP in hickory
  • Method for cloning promoter sequence of CcPIP in hickory

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Embodiment Construction

[0070] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0071] The sequence of the CcPIP promoter in the pecan obtained in the present invention is shown in SEQ ID: 1.

[0072] Analysis after sequencing showed that 876bp sequence of hickory CcPIP1 gene promoter was obtained. The results of analysis of the basic promoter region and regulatory elements by using software such as PlantCARE and PLACE showed that this sequence has the basic transcriptional elements of the promoter: 2 CAAT boxes, located at -601 and -348, are in the sequence of the promoter and the enhancer. Regulatory elements, responding to ABA and drought response; 7 TATAboxes, located at -135, -418, -435, -493, -507, -850, -864, are promoter central elements; 1 G-box, located at - 783, in response to anaerobic, light, elicitor, ABA and MeJA treatments, a MYC (CANNTG) element and a MYB (WAACCA) element locate...

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Abstract

The invention discloses a method for cloning the promoter sequence of CcPIP in hickory. The method comprises the following steps: extracting DNA in hickory leaf; designing and synthesizing a primer, and carrying out joint treatment; connecting a joint with an enzyme digestion fragment; carrying out PCR amplification, and recovering the above obtained PCR product; connecting the recovered product with a PMD18-T vector; and determining and analyzing the sequence. The method has the characteristics of cost saving, simplicity and feasibility, and is suitable for the wide application.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a method for cloning the CcPIP promoter sequence in pecan. Background technique [0002] Hickory nut (Carya cathayensis Sarg.) belongs to the genus Hickory in the family Juglandaceae. Hickory nut oil is fragrant and nutritious. It is an excellent edible oil. Hickory is mainly distributed around the Tianmu Mountains at the junction of Zhejiang and Anhui provinces, located at 29°-31° north latitude and 118°-120° east longitude, including Lin'an, Chun'an, Anji, Jianguo in Zhejiang and Ningguo, Shexian, Jinguo in Anhui De, Jixi and other counties and cities. Hickory is an important economic pillar for farmers in the mountainous areas of northwestern Zhejiang to get rid of poverty and become rich, and it is an important ecological and economic forest tree species. [0003] However, hickory has a long vegetative growth time, and the tree is tall, difficult to pick, and it is dif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/10
Inventor 郑炳松沈忆珂何勇清方佳方仲相江波司马晓娇任韡王腾飞
Owner ZHEJIANG FORESTRY UNIVERSITY
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