In the present study, we designed and constructed a chimeric multi-epitope gene of ALV-J to develop a potential multi-epitope vaccine using a reverse vaccinology approach.
Total RNA was extracted using Trizol reagent and reverse transcribed using a Reverse Transcription System (Promega).
Reverse transcription (RT) was carried out by using a reverse transcription kit (Applied Biosystem).
RNA was then reverse transcribed using a Reverse Transcription System (Promega Co., Madison, WI, USA).
Reverse transcription was performed using a reverse complement DNA-oligonucleotide to the linker sequence.
First-strand complementary DNA was reverse-transcribed using a Reverse Transcriptase MIX Kit (Toyobo, Osaka, Japan).
The RNA (1 mg) was reversed to cDNA by using a Reverse Transcription Kit (Takara).
The propensities were then fit to continuous curves and converted to a potential using a reverse-Boltzman relationship.
The cDNA encoding for α-1,3-arabinofuranosidase was reverse by reverse-transcriptase-PCR using a reverse-transcriptase (Cat, No, M531A, Promega) and oligo-dT primer.
This evidences the presence of a reverse potential barrier for ML disappearance.
