关键词: 蜡梅, cDNA文库, 脂转移蛋白基因, RT-PCR

Abstract: 【OBJECTIVE】To construct the cDNA library of Chimonanthus Praecox flower and analyze the expression of LTP cDNA ; 【METHOD】The cDNA library was constructed from flowers of Chimonanthus Praecox , using the SMART cDNA Library Construction Kit. The expression of LTP cDNA was analysed through reverse transcription polymerase chain reaction (RT-PCR);【RESULTS】The primary library has a high titer of 1.4×106 pfu/ml, in which 98% clones are recombinant and the insert cDNAs are more than 0.5 kb, with an average of 1.1 kb. The amplified library has a titer of 1010pfu/ml. The positive signals of PI, AP3 and LTP gene were detected by PCR in the amplified library, though they were expressed at different abundances. The LTP gene from the amplified library has no intron, which showed no genomic DNA in the library. RT-PCR showed LTP gene had high transcript levels during the different blooming stages, which is consistent with the fact that it was expressed at high abundance in the mini EST library;【CONCLUSION】This high quality cDNA library can be further used for gene clone and ESTs analysis and provides a useful tool for the study of the molecular mechanisms of winter-blooming of Chimonanthus Praecox.

Key words: Chimonanthus Praecox, cDNA Library, LTP, RT-PCR