Identification of the core promoter of the pmel gene of Bashang long-tail chickens / 生物工程学报

To explore the activity of the pmel core promoter of Bashang long-tail chickens, we constructed dual-luciferase expression vectors and transiently transfected into DF1 cells with Lipofectamine 2000. We measured the luciferase activity with the dual-luciferase detection kit. The 1 268 bp fragment in 5-flanking region of the pmel gene in Bashang long-tail chickens was cloned. The region from -1 200 bp to +68 bp included 2 CpG islands and multiple transcription factor binding sites. We constructed 9 expression vectors with different promoter regions and a mutant vector of the core promoter region of the pmel gene of Bashang long-tail chickens. The core promoter region from -840 bp to +68 bp was identified in the pmel gene. The region from -590 to -525 bp negatively regulated the pmel gene during the transcription process. The -840--590 bp and -525--266 bp regions were positive regulatory regions. The polymorphic sites (-456, -435, -410, -374 and -341) had a significant effect on the promoter activity of the pmel gene.

Cloning, Sequencing and Expression of Trichinella spiralis p49 Gene / 中国寄生虫学与寄生虫病杂志

Objective To conduct cloning, sequencing and expression of Trichinella spiralis ES antigen p49 gene. Methods RT-PCR was used to amplify the specific gene fragment from the total RNA of Trichinella spirais larvae. The PCR product was ligated to the T-vector and the recombinant plasmid was verified by sequencing. T-p49 and pGE-4T-3 were treated by both BamHI and XhoI. The ligation reaction was catalyzed by T4 DNA ligase. Results The p49 gene was cloned by using RT-PCR. Sequence analysis showed that the p49 gene obtained was consistent to the p49 sequence reported in the database. The expressed protein was shown as a new band at SDS-PAGE. BLAST analysis demonstrated that this p49 gene was 99% identical to the p49 gene reported and to the 43 kDa secreted glycoprotein gene in the database. Conclusion p49 gene from Trichinella spiralis larvae was cloned, sequenced and expressed.