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 sing le-tube (ST) RT-PCR without interfering with the presence of genomic D NA of cell was identified. The primer detected TNFR-I from 1 ng of cel lular RNA by ST RT-PCR, and was not crossreacted with TNFR-II despite of the homologous sequence between TNFR-I and -II. In addition, the me chanism of the RNA specificity of the primer was investigated. An intr on 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 str ucture. Thus, it appears that the RNA specificity of the primer might be resulted from the antisense primer hybridizing to exon-intron junct ion. 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.