Degenerative primer design and gene sequencing validation for select turkey genes

作者：Hutsko Stephanie L; Lilburn Michael S^*; Wick Macdonald

来源：Poultry Science, 2016, 95(6): 1403-1405.

DOI：10.3382/ps/pew009

• transcription
• sequencing
• cloning
• homology
• primers

We successfully designed and validated degenerative primers for turkey genes MUC2, RPS13, TBP and TFF2 based on chicken sequences in order to use gene transcription analysis to evaluate (quantify) the mucin transcription to probiotic supplementation in turkeys. Primers were designed for the genes MUC2, TFF2, RPS13 and TBP using a degenerative primer design method based on the available Gallus gallus sequences. All primer sets, which produced a single PCR amplicon of the expected sizes, were cloned into the TOPOR (R) vector and then transformed into TOP 10 (R) competent cells. Plasmid DNA isolation was performed on the TOP10 (R) cell culture and sent for sequencing. Sequences were analyzed using NCBI BLAST. All genes sequenced had over 90% homology with both the chicken and predicted turkey sequences. The sequences were used to design new 100% homologous primer sets for the genes of interest.

• 出版日期2016-6