Construction of cDNA Library of HPT-8 Cells Infected with Brucella menlitensis and Analysis of EST Sequence

Abstract:

The cDNA library of HPT-8 cells infected with Brucella melitensis 027 strain was constructed in this study. Total RNA were extracted from HPT-8 cells at 20 min, 1 h, 2 h, 3 h and 4 h post infection respectively. With cDNA synthesized by reverse transcriptase, homologous recombination construct infected HPT-8 cell cDNA library. The partial genes were sequenced and classifid by function. The capacity of cDNA library was 1.43 × 106, the recombinant rate was 96.92 percent. The size of the inserted fragments is between 0.2-5 kb. 63 clones of cDNA library were random selected and sequenced. The rates of transcription, energy metabolism, transporter, cytokine are relatively high. The results showed that cDNA library was successfully constructed. The cDNA library of HPT-8 cells infected with Brucella melitensis 027 strain may facilitate to identify the receptors associated with the resistance against Brucella in host cells and to cast new light on the mechanism of cellular tropism.