[A modified TAIL-PCR and its application in isolating gene promoter of wheat].
Sheng Wu Gong Cheng Xue Bao
; 24(4): 695-9, 2008 Apr.
Article
em Zh
| MEDLINE
| ID: mdl-18616185
ABSTRACT
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.
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Base de dados:
MEDLINE
Assunto principal:
Triticum
/
Reação em Cadeia da Polimerase
/
Regiões Promotoras Genéticas
Idioma:
Zh
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
China