|Table of Contents|

Molecular Cloning and RNAi Expression Vector Construction of MADS-box Gene from Castanea mollissima (PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2014年24期
Page:
92-98
Research Field:
Publishing date:

Info

Title:
Molecular Cloning and RNAi Expression Vector Construction of MADS-box Gene from Castanea mollissima
Author(s):
LI Lin-ling12LIAO Zhi-qin12CHEN Xiao-ling12CHENG Shui-yuan13CHENG Hua12
1.Economic Forest Germplasm Improvement and Comprehensive Utilization of Resources of Hubei Key Laboratories,Huanggang,Hubei 438000;
2.College of Life Science,Huanggang Normal University,Huanggang,Hubei 438000;
3.School of Biology and Pharmaceutical Engineering,Wuhan Polytechnic University,Wuhan,Hubei 430023
Keywords:
Castanea mollissimaMADS-box geneflowering periodmaturation stage
PACS:
S 664.203.6
DOI:
-
Abstract:
Taking the flowers and leaves of chestnut ‘Meiguihong’ as materials,the cDNA sequence of MADS gene was isolated from the flowers and leaves according to RACE technology and EST database from NCBI,plant expression vector of RNA interference pCl301-ubi-CmMADS-dsRNAi had also constructed by the MADS ene to study effect of blossom key gene on flower bud differentiation of Chinese chestnut.The results showed that,CmMADS was 922 bp containing an open reading frame(ORF) of 681 bp,which encoded 227 amino acids with a predicted molecular mass of 25.87 kDa and the theoretical isoelectric point(PI) of 6.27,CmMADS was an intro-free gene,and its deduced polypeptide contained a M box of 58 amino acids in the N terminal.The secondary structure of CmMADS was mainly composed of alpha helix and random coil.Comparative analysis showed that CmMADS had a high similarity to other plant MADS proteins,and contained all the M box and K box.The homology based structural modeling showed that CmMADS had the typical structure of Malus domestica MADS.Phylogenetic tree revealed that CmMADS and TrMADS3 were assigned to the same clade.Use BanH I and Kpn I and enzyme intermediate carrier pBluescript SK plus-FR and plant expression vector pcl301-ubi,recycling pBluescript SK plus- FR enzyme cut into small pieces,connected pcl301-ubi in larger pieces,a plant expression vector pCl301-ubi-CmMADS-dsRNAi.Next build good carrier of RNA interference and conversion of agrobacterium mediated by its putting the recombinant plasmid into tobacco,the function of the carrier for further study of the interference and CmMADS gene function.

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Last Update: 2014-12-24