Restricted expression of new HERV-K members in human teratocarcinoma cells

Northern hybridization with five HERV-K family members, previously cloned from human teratocarcinoma genomic DNA, indicated that two (HERV-(K)27 and -(K)67) of the five clones are expressed in these teratocarcinoma cells. These two clones are closely related (98.49%), however, and Northern blot hybridization lacks the specificity to distinguish between their respective mRNAs. Therefore, PCR analysis with mixed oligonucleotide primers homologous to conserved retroviral pol gene regions was employed to amplify cDNA synthesized from teratocarcinoma cell RNA. This amplification scheme yielded two novel HERV-K family members, HERV-(K)55 and HERV-(K)91. Clone HERV-(K)55 has approximately 98% nucleotide sequence identity to clones HERV-(K)27 and -(K)67. Subsequent RNase protection assays confirmed the expression of HERV-(K)55 and indicated that clones HERV-(K)27 and -(K)67 were not expressed in these cells. One interpretation is that the HERV-(K)27 and -(K)67 probes detected transcripts of clone HERV-(K)55 or other closely-related elements because of their high homologies. In addition, clone HERV-(K)91, which has approximately 81% nucleotide sequence identity to clones HERV-(K)27, -(K)67, and -(K)55, was obtained only from teratocarcinoma 2102E-Pr cells, but the RNase protection assay showed that this clone is also expressed in other human teratocarcinoma cell lines.