[Construction of expression vector with BG2 gene and its transformation in wheat].
beta-1,3-glucanase(BG2) is one of the pathogensis-related-proteins(PR). Study of these proteins and their related genes is one of the hot points in plant genetic engineering of disease resistance for a long time. In this research, specific primers were designed with the enzyme cleavage site of Spe I in its forward one and Not I site in the backward according to the BG2 gene sequence. Using this pair of primers, BG2 gene, which was contained in the plasmid of pRTL2, was amplified and confirmed by sequencing the amplified fragment inserted into T-easy vector. The positive clone containing BG2 gene was digested with the enzymes of Spe I/Not I and then BG2 gene was inserted into the Xba I/Not I sites of super expression binary vector pATC940. The reconstructed expression vector named as pATCBG2 was introduced into the wheat of Longfumai10 and Longfumai3 (Triticum aestivum L. em. Thell) through the particle gun transformation method. The Kanamysin resistant (Km') transformants were obtained. PCR, Dot-blotting and PCR-Southern hybridization analysis showed that the BG2 gene was integrated into the genome of wheat. Result of pathogen inoculation assay on the transgenic plants showed that the transgenic plants had a higher resistant disease score of 1-2 grade than the control.
Xing
Quan-Hua
QH
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Wang
Guang-Jin
GJ
Shi
Jin-Feng
JF
Wang
Yue-Guang
YG
Li
Zhong-Jie
ZJ
Liang
Feng-Shan
FS
Jin
De-Min
DM
Wang
Bin
B
D001203:Ascomycota / Q000254:growth & development; D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods; D018744:DNA, Plant / Q000235:genetics / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D043326:Glucan 1,3-beta-Glucosidase / Q000235:genetics*; D007113:Immunity, Innate / Q000235:genetics; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics* / Q000382:microbiology
D018744:DNA, Plant; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; D043326:Glucan 1,3-beta-Glucosidase; C117648:beta-1,3-exoglucanase