Construction of a Subtracted cDNA Library of Hyperlipidemia-sensitive Rabbit Liver by Suppression Subtractive Hybridization

Objective: To Construct a subtracted cDNA library of hyperlipidemia sensitive rabbits′liver by suppression subtractive hybridization(SSH).Methods:The mRNAs were extracted from hyperlipidemia sensitive and hyperlipidemia insensitive rabbit liver respectively and converted into double strand cDNAs. After digestion with restriction enzyme, the cDNA of hyperlipidemia sensitive group was subdivided into two portions and each one was ligated with different adaptors. The differentially expressed cDNA were obtained by two rounds of both hybridization and suppression PCR.The PCR products were inserted into T/A vector to set up the subtractive cDNA library.The clones selecting were amplified by PCR and identified. Results:The amplified library contained 500 positive bacteria clones, including 463 clones which had an inserts from 250 to 700 bp PCR analysis. Conclusion: A subtracted cDNA library of differentially expressed genes in hyperlipidemia sensitive rabbit liver was constructed successfully by SSH and T/A cloning techniques.