[Cloning of y3 gene encoding a tobacco mosaic virus inhibitor from Coprinus comatus and transformation to Nicotiana tabacum].
OBJECTIVE
The protein Y3 was a TMV inhibitor which was encoded by y3 gene. The aim of this work was to clone the full length of y3 gene from Coprinus comatus and to reveal its inhibitory function to TMV in in vivo conditions.


METHODS
We amplified the unknown 5'- terminal cDNA sequence of y3 gene with 5'- Full RACE Core Set (TaKaRa), obtained the full length of this gene by RT-PCR, constructed the expression plasmid pCAMBIA1301-y3 via inserting gene y3 sequence, CaMV 35 S promoter, and NOS terminator at MCS and transformed it into Nicotiana tabacum via agrobacterium-mediation.


RESULTS
The full length of y3 gene was 534 bps including one ORF encoding 130 amino acid residues (GenBank Accession No. GQ859168; EMBL FN546262). The cDNA sequence and its deduced amino acid sequence showed high similarity (94%) to the published fragment of y3 gene sequence. Northern blot analysis proved the transcription of y3 gene in transgenic tobacco plants. The transgenic plants inoculated with TMV expressed the inhibitory activity to TMV.


CONCLUSIONS
We cloned the full length of y3 gene and obtained transgenic tobacco plants. The expression of y3 gene in transgenic plants improved the inhibitory activity to TMV. The cloning and expression analysis of y3 gene might provide background information for future studying of y3 gene.
Wang
Xueren
X
The Provincial Key Laboratory of Biotechnology, School of Life Science, Northwest University, Xi'an 710069, China. wangxueren56@126.com
He
Tao
T
Zhang
Gaina
G
Hao
Jianguo
J
Jia
Jingfen
J
D000363:Agaricales / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000276:immunology / Q000382:microbiology; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology / Q000821:virology; D014026:Nicotiana / Q000235:genetics* / Q000276:immunology / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000502:physiology*; D014170:Transformation, Genetic
D005656:Fungal Proteins