1.A modified TAIL-PCR and its application in isolating gene promoter of wheat.
Yanguang QIU ; Jinghan TIAN ; Rongchao GE ; Baocun ZHAO ; Yinzhu SHEN ; Zhanjing HUANG
Chinese Journal of Biotechnology 2008;24(4):695-699
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.
Base Sequence
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Genes, Plant
;
genetics
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Molecular Sequence Data
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Polymerase Chain Reaction
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methods
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Promoter Regions, Genetic
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genetics
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Triticum
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genetics
;
metabolism