[A modified TAIL-PCR and its application in isolating gene promoter of wheat].
Using a modified TAIL-PCR technique, the 5' -flanking region of the X gene in wheat was successfully isolated. Two novel modifications of the TAIL-PCR were introduced here: using a battery of random 10-mers as the short arbitrary primers instead of three degenerate 16-mers; using 29 degrees C instead of 44 degrees C as the annealing temperature for the low-stringency cycle; increasing five high-stringency cycles and reducing five low-stringency cycles; and using single primers for the third round of product identification. Isolated 5' -flanking region was fused to the GUS gene, and tested for expression in Arabidopsis plants. Histochemical analysis of the transgenic plants showed the report gene was driven by isolated 5'-flanking region. Modified TAIL-PCR technique could isolate rapidly the promoter of any gene from organisms with large genomes.
Qiu
Yanguang
Y
College of Life Sciences, Hebei Normal University, Shijiazhuang 050016, China.
Tian
Jinghan
J
Ge
Rongchao
R
Zhao
Baocun
B
Shen
Yinzhu
Y
Huang
Zhanjing
Z
D001483:Base Sequence; D017343:Genes, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014908:Triticum / Q000235:genetics* / Q000378:metabolism