The Construction Of Full-length CDNA Library From Saussurea Involucrata Kar.et Kir In Xinjiang And Its Sequence Analysis

The full-length cDNA library of Saussurea involucrata Kar.et was constructed by using SMART(switching mechanism at 5＇end of RNA transcript) technology .The total RNA was extracted and was generated full-length cDNA, The PCR product were digested by proteinase K and SfiI ,and fractionated with the Chroma spin -400 Column.The cDNAs of more than 400bp were collected and ligated intoλTripLEx2 vector. The λ-phage packaging reaction was used for the final construction of the cDNA library. The titer of unamplified cDNA library consisted of 4.03×107 independent clones with a recombinants rate of 93%.While the amplified library 1.75×109 pfu/ml. 120 positive clones were randomly selected for plasmid DNA extraction and sequencing. 80 EST sequences were obtained. After BLAST identity analysis, 42 genes of known function and 10 genes of unknown function were obtained.