Identification and characterization of an RNA specific primer for human tumor necrosis factor receptor I (TNFR-I)

An RNA specific primer amplified TNFR-I cDNA from cellular RNA by single-tube (ST) RT-PCR without interfering with the presence of genomic DNA of cell was identified. The primer detected TNFR-I from 1 ng of cellular RNA by ST RT-PCR, and was not cross-reacted with TNFR-II despite of the homologous sequence between TNFR-I and -II. In addition, the mechanism of the RNA specificity of the primer was investigated. An intron of 0.43 kb was inserted in the 5'-925 of TNFR-I mRNA sequence. The result corresponded with the previous determination of TNFR-I gene structure. Thus, it appears that the RNA specificity of the primer might be resulted from the antisense primer hybridizing to exon-intron junction. Owing to its high sensitivity and specificity to TNFR-I RNA, the RNA specific primer may be potentially utilized for the determination of human TNFR-I gene expression by in situ RT-PCR.