Carya cathayensis auxin efflux carrier protein CcPILS gene cloning and expression analysis method

A technology for outputting vectors and analysis methods, applied in the directions of biochemical equipment and methods, genetic engineering, plant genetic improvement, etc., can solve problems such as difficulty in improving quality, difficulty in grafting pecans, and difficulty in variety selection, and achieves the effect of solving and promoting

Inactive Publication Date: 2017-04-26
ZHEJIANG FORESTRY UNIVERSITY
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Problems solved by technology

[0007] The purpose of the present invention is to overcome the above-mentioned deficiencies in the prior art, and provide a method for cloning and expression analysis of the

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  • Carya cathayensis auxin efflux carrier protein CcPILS gene cloning and expression analysis method
  • Carya cathayensis auxin efflux carrier protein CcPILS gene cloning and expression analysis method
  • Carya cathayensis auxin efflux carrier protein CcPILS gene cloning and expression analysis method

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[0036] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. However, it should be understood that the specific embodiments described here are only used to explain the present invention, and are not intended to limit the scope of the present invention. Also, in the following description, descriptions of well-known structures and techniques are omitted to avoid unnecessarily obscuring the concept of the present invention.

[0037] The embodiment of the present invention provides a method for cloning and expression analysis of the hickory auxin export carrier protein CcPILS gene, comprising the following steps:

[0038] A), preparation material: plant material is hickory grafting sample, starts sampling before and after grafting at test site, gets rootstock and scion respectively as contrast before grafting, then ...

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Abstract

The invention discloses a Carya cathayensis auxin efflux carrier protein CcPILS gene cloning and expression analysis method. The method includes the steps of: material preparation, RNA extraction and purification, CcPILS gene RACE, bioinformatics analysis, and temporal and spatial expression analysis. The method provided by the invention clones Carya cathayensis PIL homologous gene CcPILS by RACE technique, the gene has a full-length sequence of 1541bp, wherein the open reading frame sequence is 1263bp, bioinformatics software analysis is applied to predict that the sequence encodes 420 amino acids, the CcPILS protein molecular weight is about 46.22KD, PI is 5.38, the protein is located at an endoplasmic reticulum membrane, is equipped with 5 transmembrane hydrophobic structural domains respectively at the N terminal and C terminal, and is separated by a middle hydrophilic loop. The gene has high homology with Arabidopsis thaliana AtPILS5 and AtPILS7, and belongs to Clade III subtribe of PILS gene family. Fluorescence quantitative RT-PCR analysis shows that before and after Carya cathayensis grafting, CcPILS has expression trend in scion consistent with that in rootstock, and provides a beneficial promoting effect on solution of the problems of difficult Carya cathayensis grafting, difficult variety breeding and difficult quality improvement, etc.

Description

[0001] 【Technical field】 [0002] The invention relates to the technical field of pecans, in particular to a method for cloning and expression analysis of the pecan auxin export carrier protein CcPILS gene. [0003] 【Background technique】 [0004] Plants regulate auxin content in different tissues through auxin metabolism and intercellular transport, and then generate signals to regulate plant tropism, the occurrence and development of lateral organs, etc. Polar transport is an important feature of auxin, which is coordinated by the auxin input carrier AUX / LAX protein family (AUXIN RESISTANT1 / LI-KE AUX1) and the export carrier PIN-FORMED (PIN) protein family. Barbez et al. found another auxin intracellular transport-related gene family with 7 members. Although the sequence similarity between this gene and PIN is very low (10%-18%), its topological structure is similar to that of PIN protein, so it is named PILS (PIN-LIKES, PILS). PILS can be divided into three subfamilies, Cl...

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Application Information

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IPC IPC(8): C12N15/29C12N15/10C12Q1/68
CPCC07K14/415C12N15/10C12Q1/6851C12Q2531/113C12Q2549/119C12Q2563/107C12Q2545/101
Inventor 郑炳松高柳肖郭文彬闫道良袁虎威赵亮徐栋斌童亚菲
Owner ZHEJIANG FORESTRY UNIVERSITY
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