Isolation and complete nucleotide sequence of the gene for bovine parathyroid hormone.

The structure of the bovine parathyroid hormone (PTH) gene has been analyzed by Southern blot hybridization of genomic DNA and by nucleotide sequence analysis of a cloned PTH gene. In the Southern analysis, several restriction enzymes produced single fragments that hybridized to PTH cDNA suggesting that there is a single bovine PTH gene. The restriction map of the cloned gene is the same as that determined by Southern blot analysis of bovine DNA. The sequence of 3154 bp of the cloned gene has been determined including 510 bp and 139 bp in the 5' and 3' flanking regions, respectively. The gene contains two introns which separate three exons that code primarily for: (i) the 5' untranslated region, (ii) the pre-sequence of preProPTH, and (iii) PTH and the 3' untranslated region. The gene contains 68% A + T and unusually long stretches of 100- to 150-bp sequences containing alternating A and T nucleotides in the 5' flanking region and intron A. The 5' flanking region contains two TATA sequences, both of which appear to be functional as determined by S1 nuclease mapping. Compared to the rat and human genes, the locations of the introns are identical but the sizes differ. Comparable human and bovine sequences in the flanking regions and introns are about 80% homologous.

Restriction of Moloney murine leukemia virus replication in Moloney murine sarcoma virus-infected cells.

MuSV349 is a TB-cell line which produces infectious MuSV with little or no MuLV detectable by the XC assay. The apparent restriction of MuLV replication in MuSV349 cells was investigated. A replication-competent helper virus, with protein composition nearly identical to that of Mo-MuLV was isolated from the viruses produced by MuSV349 cells. This helper MuLV after it was separated from MuSV and upon infection of TB cells produced viral titer similar to that of Mo-MuLV-infected TB cells indicating that its replication might have been restricted in the MuSV349 cells. To find out whether the suppression of the helper virus replication is due to the genetic peculiarities of the virus or MuSV349 cells, the relative amounts of MuLV and MuSV produced by several distinct clonal MuSV isolates (derived from a common progenitor) upon superinfection with Mo-MuLV were determined. The results of these experiments showed that while both SV7 and SV15F on coinfection with Mo-MuLV produced MuSV in excess over MuLV; and ts110 and tsSV13 on coinfection with Mo-MuLV produced MuLV in excess over MuSV. Since the same Mo-MuLV is used in these experiments and since upon transfer to a different cell, SV7, SV15F, and ts110 retain the property to restrict or not restrict MuLV replication it appears that the above property is determined by the genetics of the MuSV.

Phylogenetic origin of the chloroplast and prokaryotic nature of its ribosomal RNA.

The 16S ribosomal RNA of the Euglena gracilis chloroplast has been characterized in terms of its two-dimensional electrophoretic "fingerprint" (T1 ribonuclease). Results show it to be a typically prokaryotic 16 S rRNA. By the present criterion, different chloroplasts are shown to be related to one another and at least distantly to blue-green algae and perhaps to Bacillaceae. These results argue in favor of an endosymbiont origin of the chloroplast.