Soybean transformation based on RNA interference(RNAi)can specifically silence the homologous target genes in mRNA level,providing a new strategy for disease-resistant breeding.In this study,we made the amino acid sequence alignment of eIF4E from ten different kinds of plants which have been identified to interact with the virus VPg,then we determined the conserved interval of soybean eIF4E was 62-237 amino acid sequence and cloned the 406 bp interference fragment named eIF4Ei (including 58 bp specific recombination sequence attB).Using the GATEWAY technology,the RNAi vector pB7GWIWG2(Ⅱ)-eIF4Ei was constructed,which was identified by PCR amplification,sequencing and restriction digestion.The identification of the recombinant expression vector ensured the eIF4Ei interference fragment no mutation in the process of vector construction and two ccdB sites were all replaced by eIF4Ei,in addition,the insert direction of the two interference fragments in the terminal plasmid was conversed.Overall,the RNAi vector was successfully constructed to be an inverted repeat structure,providing the basic materials for cultivating new germplasm of soybeans resistant to SMV using the soybean transformation technology based on RNAi.
GAO Le
,
ZHAI Rui
,
DING Xue-ni
,
LI Kai
,
ZHANG Hong-yun
,
WANG Tao
,
ZHI Hai-jian
. Cloning of Interference Fragment of Eukaryotic Translation Initiation Factor 4E of Soybean and Construction of Its RNAi Vector[J]. Acta Agriculturae Boreali-Sinica, 2014
, 29(4)
: 1
-6
.
DOI: 10.7668/hbnxb.2014.04.001
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