Genetic Analysis of a Mouse t Complex Locus That Is Homologous to a Kidney Cdna Clone

A mouse kidney cDNA clone, pMK174, identifies restriction fragment length polymorphisms (RFLPs) that map to two unlinked loci. One, designated D17Rp17, has been mapped near quaking, (qk), on chromosome 17 using three sets of recombinant inbred (RI) strains. A study of several t haplotypes resulted in the identification of t-specific alleles of D17Rp17 that map to the proximal half of the t complex. Neither t-specific nor wild-type D17Rp17 alleles are present in chromosomes carrying either the T Orleans (TtOrl) or the T hairpin tail (Thp) deletions. Comparison with other molecular markers indicates that pMK174 identifies a new proximal t complex locus, Rp17. The second locus identified by pMK174, termed D4Rp18, is tentatively assigned to chromosome 4 by mouse-Chinese hamster somatic cell hybrid analysis.     

Inhibitory Effect of Autoclaving Whey-Based Medium on Propionic Acid Production by Propionibacterium shermanii

Propionic acid production by Propionibacterium shermanii was compared in pasteurized and autoclaved whey-based media. Propionic acid production decreased with increasing whey concentration in autoclaved media but not in pasteurized media. Increasing the yeast extract concentration from 5 to 10 g/liter greatly reduced the inhibitory effect of autoclaving.     

Production of High-Viscosity Whey-Glucose Broths by a Xanthomonas campestris Strain

Acclimation of a sandy soil to an air-natural gas mixture stimulated the biological oxidation of chloroform to carbon dioxide. Acetylene and methane inhibited chloroform oxidation. Chloroform oxidation continued up to 31 days in the absence of methane. Chloroform oxidation rates increased at chloroform concentrations up to 5 μg g of soil^-1.     

Walleye, Stizostedion vitreum, and northern pike, Esox lucius, populations in three Alberta lakes

Results are presented from a survey of walleye and northern pike in Ethel, Marie and Wolf Lakes, Alberta conducted May-September, 1983. Three types of experimental gill net were fished, mainly inshore. Northern pike were abundant in all three lakes but white sucker and yellow perch also were common. Walleye were caught in largest numbers in Wolf Lake. Age and growth of both walleye and northern pike were determined from examination of the opercular bones. The walleye in the three lakes were represented by a few strong year classes but there was no synchronism between the lakes. The strong 1975 year class in Ethel and Wolf Lakes may have been correlated with high spring precipitation and heavy run-off. Although there were dominant year classes in the northern pike populations (especially the 1979 year class) the variability was not so marked as in the walleye. There were no significant differences between length-at-age for year classes of walleye in Ethel and Marie or northern pike in all three lakes. In Wolf Lake the 1978 and 1979 year classes grew faster than in previous years. Walleye and northern pike growth was well described by the von Bertalanffy growth model. Females of both species grew to a greater ultimate length and had a longer lifespan than the males. Fecundity data are presented for walleye from Wolf Lake (log₁₀ absolute fecundity = 0.856 + 2.441 log₁₀ length) and northern pike from Marie Lake (log₁₀ absolute fecundity =?2.671+4.052 log₁₀ length). The stomach contents of both species were examined during May and June. The majority of walleye had empty stomachs. Walleye and northern pike in Ethel and Wolf Lakes fed on a variety of fish and invertebrate species but both fed mainly on fish (in particular whitefish) in Marie Lake. The similarity of diet suggests competition for prey species. A walleye yield to commercial fishermen of c. 0.69 kg ha^?1 year^?1 in Wolf Lake has made up 8% of the total catch. Since 1970/71 when mesh size of nets was increased, the yield has been reduced to c. 0.30 kg ha^?1 year^?1. Anglers remove an additional c. 0.48 kg ha^?1 year^?1 walleye from Wolf Lake. Northern pike yields to commercial fishermen have been c. 2.00 kg ha^?1 year^?1 in Wolf Lake. Ethel and Marie Lakes have yielded only poor catches of walleye and northern pike. A yield model was used to illustrate that faster-growing northern pike have higher potential yields than walleye. Walleye produce higher yields in Wolf Lake than in Marie Lake, the reverse being true for northern pike. It is suggested that northern pike could be cropped at a higher rate in Marie and Wolf Lakes with a possible improvement in walleye stocks.     

A restricted human antitetanus clonotype shares idiotypic cross-reactivity with tetanus antibodies from most human donors and rabbits: reactivity with antibodies of widely differing electrophoretic mobility

Antitetanus antibodies from each of 20 hyperimmunized human donors were isolated on a tetanus immunoadsorbent, eluted with acidic buffer, and examined by isoelectric focusing (IEF). There was electrophoretic restriction, as determined by IEF, in the IgG of only 20% of the purified antibodies studied. The remaining 80% showed a more diffuse polyclonal spectrotype. Several IEF bands from the most electrophoretically restricted sample were isolated and used to immunize rabbits. Virtually every IgG-IEF band in the antitetanus antibodies of the original donor shared idiotypic cross-reactivity as detected by one or more of the three rabbit anti-Id reagents, even though major qualitative differences in binding from one rabbit anti-Id reagent to another were noted. Antitetanus antibodies of each of the 20 donors were separated by IEF and transferred to a nitrocellulose membrane. By using a sensitive and specific ELISA detection method, cross-reactivity was detected with the rabbit anti-Id reagents in 1 to 50% of the antitetanus antibodies of individual donors. This cross-reactivity was greater than 10% in 15 of the 19 antisera studied. In addition, these cross-reactive antibodies had very different electrophoretic mobility. Binding of the rabbit anti-Id reagents to the tetanus antibodies was almost completely blocked by pretreatment with soluble tetanus toxoid antigen. This idiotypic cross-reactivity with antibodies of different electrophoretic mobility from the same and unrelated donors suggests sharing among these antibodies of one or more of the germ-line DNA-encoded hypervariable regions present in the antibody-combining site.     

Intracellular transport of herpes simplex virus gD occurs more rapidly in uninfected cells than in infected cells.

A mouse L cell line which expresses the herpex simplex virus type 1 immediate-early polypeptides ICP4 and ICP47 was cotransfected with a cloned copy of the BglII L fragment of herpes simplex virus type 2, which includes the gene for gD, and the plasmid pSV2neo, which contains the aminoglycosyl 3'-phosphotransferase (agpt) gene conferring resistance to the antibiotic G418. A G418-resistant transformed cell line was isolated which expressed herpes simplex virus type 2 gD at higher levels than were found in infected cells. The intracellular transport and processing of gD was compared in transformed and infected cells. In the transformed Z4/6 cells gD was rapidly processed and transported to the cell surface; in contrast, the processing and cell surface appearance of gD in infected parental Z4 cells occurred at a much slower rate, and gD accumulated in nuclear membrane to a greater extent. Thus, the movement of HSV-2 gD to the cell surface in infected cells is retarded as viral glycoproteins accumulate in the nuclear envelope, probably because they interact with other viral structural components.     

Evidence for [D-Ala2,D-Leu5]Enkephalin-Induced Supersensitivity to 5-Hydroxytryptamine in a Neurotumor × Brain Hybrid Cell Line (NCB-20)

A neuroblastoma X Chinese hamster embryonic brain explant hybrid cell line (NCB-20) expressed 5-hydroxytryptamine (5-HT1) receptors, linked to adenylate cyclase, which closely resembled 5-HT1 receptors previously characterized in central nervous tissue. However, the affinity of the receptors for 5-HT was only 150 nM compared to 5 nM in membranes prepared from cerebral cortex. The elevation of cyclic AMP levels in NCB-20 cells produced by 5-HT was found additive to that produced by cholera toxin but synergistic with that produced by either prostaglandin E1 (PGE1) or forskolin, suggesting that these latter two agents elevate cyclic AMP levels by a different mechanism than 5-HT. The elevation of cyclic AMP levels by either 5-HT or PGE1 was reversed by [D-Ala2,D-Leu5]enkephalin (DADLE), morphine, clonidine, and 3,4-dihydroxyphenylethylamine (dopamine) on a short (30 min) time scale. However, continued exposure to DADLE resulted in loss of the initial inhibitory effects of DADLE after 6 h and return of cyclic AMP levels to that seen with either 5-HT or PGE1 alone. When the DADLE exposure time was increased to 48 h, 5-HT produced a further twofold increase in cyclic AMP levels, but there was no increase in the responsiveness of the cells to PGE1 unless naloxone was added 1 h prior to treatment with PGE1. Scatchard analysis showed that the increased potency of 5-HT resulted from an increase in receptor affinity for 5-HT (from a KD of 150 +/- 20 nM to one of 20 +/- 7 nM), with a reduction in the number of apparent binding sites. The 5-HT supersensitivity observed in NCB-20 cells may be a good model for neurotransmitter interactions that produce desensitization or facilitation in the intact nervous system.     

Signals for site-specific cleavage of HSV DNA: maturation involves two separate cleavage events at sites distal to the recognition sequences

Mature Herpes Simplex Virus (HSV) genomes are cleaved from concatemeric precursors by a site-specific mechanism. These cleavage events are probably coupled to the encapsidation process. Sequences within the terminal repeat of HSV DNA are necessary for the cleavage and packaging reactions, and are also thought to be responsible for high frequency genome isomerization events. Here we present evidence to show that two viral DNA cleavage and packaging signals reside within a 250 bp subfragment of the terminal repeat, that the termini of mature viral DNA are generated by a process involving two separate DNA cleavages at sites distal to the cleavage signals, and that the sequences between these two cleavage sites are duplicated by the DNA maturation system.     

Structural analysis of a maize gene coding for glutathione-S-transferase involved in herbicide detoxification

Summary We have used the cDNA clone encoding maize glutathione-S-transferase (GST I) to isolate a genomic DNA clone containing the complete GST I gene. Nucleotide sequence analysis of the cDNA and genomic clones has yielded a complete amino acid sequence for maize GST I and provided the exon-intron map of its gene. The mRNA homologous sequences in the maize GST I gene consist of a 107 bp 5 untranslated region, a 642 bp coding region and 340 bp of the 3 untranslated region. They are divided into three exons by two introns which interrupt the coding region. The 5 untranslated spacer contains an unusual sequence of pentamer AGAGG repeated seven times. The inbred maize line (Missouri 17) contains a single gene for GST I, whereas the hybrid line (3780A) contains two genes. Nucleotide sequence analysis of the primer extended cDNA products reveals that the 5 untranslated regions of the two genes in the hybrid 3780A are identical except for a 6 bp internal deletion (or insertion). The amino acid sequence of maize GST I shares no apparent sequence homology with the published sequences of animal GST's and represents the first published sequence of a plant GST. re]19850813 ac]19851126