首页|Construction of yeast two-hybrid cDNA libraries for wheat nearisogenic line TcLr19 under the stress of Puccinia recondita and its preliminary appreciation

Construction of yeast two-hybrid cDNA libraries for wheat nearisogenic line TcLr19 under the stress of Puccinia recondita and its preliminary appreciation

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Two cDNA libraries for wheat near-isogenic line TcLr19 under Puccinia recondita stress were constructed by using SMART technique and homologous reorganization method.Wheat near-isogenic line TcLr19 was infected with leaf rust race 366,and total RNA was extracted from the leaves after infection for 4,8,and 12 h.The total RNAs were reverse transcribed to cDNA by using oligo(dT) primer and random primer,respectively.According to the evaluation on quality,the transformation efficiency was about 1.32 × 106 and 1.0 × 106 transformants/3 μg pGADT7-Rec,respectively,and the library titers were up to 2.62 × 108 and 3.51 × 108 pfu/mL,with 93% and 100% recombinant rate,which indicated the high quality of the two libraries for next screening.

wheat leaf rustSMART techniquehomologous reorganizationcDNA library

Lifeng ZHANG、Hui ZHOU、Fengju WEI、Ziyi CHENG、Aihua YAN、Dongmei WANG

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College of Life Science, Agricultural University of Hebei, Baoding 071001, China

2011

中国农学前沿
高等教育出版社

中国农学前沿

ISSN:1673-7334
年,卷(期):2011.5(4)
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