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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2009, Vol. 3 Issue (1) : 55-59     DOI: 10.1007/s11703-009-0017-y
RESEARCH ARTICLE |
Cloning of strawberry FaEtr2 gene and its plant expression vector construction for antisense RNA
Chunli SONG1, Pingping ZHOU1, Junlian MA2, Xia TANG2, Zide ZHANG2(), Zhixia HOU3
1. College of Chinese Veterinary,Agricultural University of Hebei,Dingzhou 073000,China; 2. College of Food Science and Technology,Agricultural University of Hebei,Baoding 071001,China; 3. The Key Laboratory for Silviculture and Conservation of Ministry of Education, Beijing Forestry University, Beijing 100083, China
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Abstract  

An ethylene receptor FaEtr2 gene was amplified by Polymerase Chain Reaction (PCR) from ripening strawberry fruit. A 1049-bp PCR product (All Star-Etr2) was cloned. Sequence analysis showed that the All Star-Etr2 nucleotide sequence had 100% identity with Chandler-Etr2 from the GenBank. A pair of primers containing restriction enzyme sites were designed and used to amplify the sequenced plasmid. The PCR product was digested by the corresponding restricted enzymes and inserted between the CaMV 35S promoter and NOS terminator of expression vector pBI121 directionally. The constructed expression vector was transformed into Agrobacterium fumefeciens LBA4404 in the follow-up research to silence a ripening-related ethylene receptor FaEtr2 gene in strawberry fruits.

Keywords strawberry      ethylene receptor      FaEtr2 gene      plant expression vector     
Corresponding Authors: ZHANG Zide,Email:zhangzde@heinfo.net   
Issue Date: 05 March 2009
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-009-0017-y     OR     http://academic.hep.com.cn/fag/EN/Y2009/V3/I1/55
Fig.1  Agarose gel electrophoretogram of PCR products
Note: M: 100 bp DNA Ladder-3K marker; 1—5: PCR products; CK: negative comparison.
Fig.2  Electrophoresis pattern of recombinant plasmid digested by R I
Note: M: 1 kb ladder marker; 1, 2: recombinant plasmid digested by R I.
Fig.3  The nucleotide sequence of all star strawberry
Note: Underlined sequences were the primer sequences.
Fig.4  Agrose gel electrophoretogram of PCR products
Note: M: 100 bp ladder marker; 1, 2: PCR products of antisense; CK: negative comparison of PCR products.
Fig.5  Agrose gel electrophoretogram of antisense PCR digested by enzymatic
Note: M: 100 bp ladder marker; 1: antisense PCR digested by I + H I; 2: PCR products of antisense.
Fig.6  Agrose gel electrophoretogram of pBI121 plasmid digested by I and H I
Note: M: 1 kb ladder marker; 1: pBI121 plasmid digested by I and H I; 2: pBI121 plasmid.
Fig.7  Detection of the antisense expression vector by enzymatic digestion
Note: 1: I/R I; 2: I/H I; 3: d III/ I; 4: d III/ H I; 5: pBI-Anti-Etr2; M: 1 kb ladder marker.
Fig.8  Agrose gel electrophoretogram of antisense expression vector transformed LBA4404
Note: M: 100 bp ladder marker; CK: comparison of negative contral; 1—3: PCR products of bacterium liquid; CK: PCR product of the plasmid.
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