Construction of subtracted cDNA library of seasickness susceptibile rat brain stem

  Objective   Isolation of genes related to seasickness susceptibility in rat's brain stem with suppression subtractive hybridization was helpful in establishing a method of predicting people seasickness suscept ibility.

  Methods   the mRNA were extracted from simulated seasickness susceptible and insusceptible rats brain stem.Suppr ession subtractive hybridization (SSH)method was used for isolating differentially expressed cDNA.After two times of suppression PCR, the cDNA fragments were inserted into T-vectors to set up the subtracted library, amplification of the library was carried out with transformation of E.coli by high votage electroperfor mation.The clones were amplified by PCR and identified.

  Results   the subtracted library contained 470 clones(260 high expressed, 210 low expressed)including 442 clones containing 250-650 bp inserts.

  Conclusion   the subtracted cDNA library of simulated seasickness susceptible rat's br ain stem was successfully constructed.It was helpful in the study of new and specific genes expressed in seasickness.