In vitro construction of a recombinant adenovirus Ad2:Ad5

A hybrid virus containing the left half of the Ad5 genome and the right half of the Ad2 genome has been constructed by ligating together in vitro the BamHI.-A fragment of Ad5 (map co-ordinates 0–59.5) to the-SawHI-A fragment of Ad2 (map coordinates 59.5–100), and using this DNA to transfect susceptible cells. Viable progeny virus has been obtained which grows as well as the parental virus without any requirement for helper virus, and probably contains a hybrid hexon polypeptide consisting of the major part of the Ad5 hexon with an Ad2 carboxy terminus.     

Murine leukemia cell hybrids: The quantity of TL antigens expressed by parental and hybrid cells fails to correlate with their sensitivity to TL antibody and complement

The quantity of thymus-leukemia (TL) antigens expressed by murine leukemia cells is significantly greater than that expressed by somatic hybrids of such cells. Based upon the results of ¹²⁵I-lactoperoxidase labeling and antibody absorption procedures, and corrected for size differences between the two cell types, the quantity of TL antigens expressed by RADA-1 cells, a radiation-induced murine leukemia cell line of strain A/J mice, is approximately 5.0 times greater than that of somatic hybrids of RADA-1 and LM(TK)^? cells. LM(TK)^? cells are a thymidine kinase-deficient TL(-) mouse fibroblast cell line. The quantity of TL antigens expressed is related only in part to their susceptibility to lysis by TL antibodies and guinea pig complement (GPC). RADA-1 cells resist lysis. The quantity of TL antigens expressed by RADA-1 cells is analogous to that formed by nonneoplastic thymocytes obtained from F₁ hybrids of two strains of TL(+) and TL(-) mice; cells from both strains are sensitive to TL antiserum and GPC. ASL-1 cells, a spontaneously occurring leukemia cell line of A/J mice, express TL antigens in significantly higher quantities than any of the cell types examined. Exposed to TL antisera, the quantity of TL antigens of ASL-1 cells, but not that of hybrid cells, gradually diminishes. ASL-1 cells convert over a 6-h period of exposure to antibody and guinea pig complement (GPC) resistance; hybrid cells remain sensitive. However, ASL-1 cells converted to TL antibody and GPC resistance continue for a time to express TL antigens in quantities similar to that of sensitive F₁ thymocytes and resistant RADA-1 cells. RADA-1 X LM(TK)^? hybrid cells, which are sensitive to TL antibodies and GPC, express the lowest quantities of TL antigens of any of the cell types examined. It is likely that differences in the quantities of TL antigens expressed by different cell lines reflect genetic mechanisms controlling TL antigen expression. The failure of TL antisera to affect the quantities of TL antigens expressed by hybrid cells is taken as an indication that genetic controls governing antigen expression may be distinguished from those involved in regulating responsiveness to specific antiserum.     

Gene fusion vectors based on the gene for staphylococcal protein A

Two plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusion, have been constructed. These vectors will allow fusion of any gene to the protein A gene, thus giving hybrid proteins which can be purified, in a one-step procedure, by IgG affinity chromatography. As an example of the practical use of such vectors, the protein A gene has been fused to the lacZ gene of Escherichia coli. E. coli strains containing such plasmids produce hybrid proteins with both IgG binding and β-galactosidase activities. The hybrid protein(s) can be immobilized on IgG-Sepharose by its protein A moiety with high efficiency without losing its enzymatic activity and they can be eluted from the column by competitive elution with pure protein A. The fused protein(s) also binds to IgG-coated microtiter wells which means that the in vivo product can be used as an enzyme conjugate in ELISA tests.     

红金麻——木槿属一个新杂种

<正> 红金麻 新杂种 Hibiscus×hongjinma Y. T. Chang, hybr. nov. H. cannabinus L. ♀× H. radiatus Cav.♂ Hybrida hortensis ( 2n=30+6sat) θ Hibisco cannabino L. ( 2n=32+4sat) et H. ranndiato Cav. (2n=70+2sat) exorta, Habitu illo simil, characteribus ad hoc magis accedit; ab illo differt floribus purpureis, folio prope basin 3—5     

Rat immunoglobulin delta heavy chain gene: nucleotide sequence derived from cloned cDNA

Rat immunoglobulin δ heavy-chain mRNA has been isolated. RNA blot analysis revealed that this mRNA with a length of 1.8 kb encodes for the secreted form of IgD. The corresponding cDNA was cloned in plasmid pBR322 and its sequence was determined. The hybrid plasmid contains a 775-bp insert comprising a partial Cδ₁ sequence and complete CδH, Cδ3, CδDC and 3' untranslated sequences. Rat and mouse IgD amino acid sequences show striking homology in Cδ3 and CδDC regions.     

Toxic ligand conjugates as tools in the study of receptor-ligand interactions

We have constructed hybrid proteins in which the toxic A chains of ricin or diptheria toxin have been linked to either asialofetuin, fetuin, or epidermal growth factor (EGF). Both ASF-RTA and ASF-DTA are potent toxins on cultured rat hepatocytes, cells that display the asialoglycoprotein receptor. Toxicity of these two compounds is restricted to hepatocytes and can be blocked by asialoglycoproteins but not the native glycoproteins or asialoagalactoglycoprotein derivatives, indicating that the toxicity of the conjugates is mediated by the hepatic asialoglycoprotein receptor. The EGF-RTA conjugate is an extremely potent toxin on cells that can bind the hormone, but is only poorly effective on cells that are unable to bind EGF. The EGF-DTA conjugate, in contrast, is unable to kill 3T3 cells and is at least two orders of magnitude less effective than EGF-RTA on A431 cells, a cell line with 1-2 X 10(6) EGF receptors per cell. However, when EGF-RTA and EGF-DTA were tested on primary liver hepatocyte cultures, which were susceptible to both ASF-RTA and ASF-DTA, both EGF conjugates were potent toxins. Sensitivity of the hepatocyte cultures to ricin toxicity increases slightly during a 52-hr culture period. In contrast, sensitivity to EGF-RTA and ASF-RTA decline dramatically during this period. Receptors for both ligands remain plentiful on the cell surface during this time.     

Dermorphins, Opioid Peptides from Amphibian Skin, Act on Opioid Receptors of Mouse Neuroblastoma x Rat Glioma Hybrid Cells

Dermorphin and its Hyp6 analogue are opiate-like heptapeptides originally discovered in frog skin and characterized by the presence of a D-Ala2 residue in their sequence. They were assayed for their capacity to compete with [3H]Leu-enkephalin for binding to opioid receptors in membranes of neuroblastoma x glioma hybrid cells. In the presence of 7 nM-[3H]Leu-enkephalin, the concentrations at which they caused 50% inhibition of [3H]enkephalin binding (IC50 values) are 0.1 micro M and 0.3 micro M, respectively. In contrast, the synthetic L-Ala2-dermorphin shows very low affinity for the opioid receptors. In addition, like other opioid peptides, dermorphin and hyp6-dermorphin inhibit the elevation by prostaglandin E1 (PGE1) of the level of adenosine 3':5'-cyclic monophosphate (cyclic AMP) (IC50 values 0.2 micro M and 0.4 micro M, respectively). The inhibition is prevented by the opiate antagonist naloxone, L-Ala2-dermorphin is at least three orders of magnitude less potent in inhibiting the PGE1-evoked increase in the level of cyclic AMP. The results show that peptides with an amino acid sequence quite different from that of the enkephalins can bind to opioid receptors of the hybrid cells.     

Hybridization between tetraploid species of Alopecurus L. (Poaceae): chromosome behaviour of natural and artificial hybrids

The meiotic behaviour of plants from mixed populations of Alopecurus pratensis × A. geniculatus and of A. pratensis × A. arundinaceus ( 2n = 4×(= 28) is reported together with observations on some artificially produced hybrids. This meiotic behaviour is correlated with the degree of hybridity as shown by the hybrid index values of the plant. In A. pratensis × A. geniculatus hybrids there are marked differences between populations in the degree of meiotic disturbance and in one population there was an almost complete breakdown of meiosis. Alopecurus pratensis × A. arundinaceus hybrids show fairly regular meiotic pairing but in most plants there are a few univalent chromosomes at metaphase I. Artificially produced hybrids of A. pratensis × A. aequalis gave meiotic configurations that suggest that the genomes of the parent species are very similar. Taken together with the results from artificially produced hybrids of A. pratensis × A. geniculatus , it is suggested that bivalent-promoting mechanisms reduce multivalent formation both in the species and the hybrids. The interaction of different genotypes involved in the control of meiosis may account for the variation in meiotic behaviour in the different populations of hybrids. Pollen fertility is reduced in most populations of hybrids and likely to be an important factor permitting introgressive hybridization.     

Soaking injury and its reversal with polyethylene glycol in relation to respiratory metabolism in high and low vigor soybean seeds

Germination and growth of unaged (high vigor) and accelerated aged (41°C and 100% RH for 4 days; low vigor) soybean [ Glycine max (L.) Merr. cv. Essex] seeds were severely reduced by soaking the seeds under water at 25°C during the first 1 to 8 h of imbibition. Soaking in 30% polyethylene glycol (PEG) did not injure high vigor seeds and improved the performance of low vigor seeds. Soaking in water increased subsequent RQ values and acetaldehyde and ethanol levels. In high vigor seeds, the increase in RQ lasted only 1 h after transfer to aerobic conditions, whereas in low vigor seeds the increase persisted into the second h. Increases in ethanol and acetaldehyde after soaking were more pronounced in low than in high vigor seeds. The data indicate that relatively short periods under water can cause water uptake injury to imbibing soybean seeds and that this injury can be avoided by osmotically reducing the initial rate of water uptake. Seeds whose vigor has declined during accelerated aging show an additional injury which increases with soaking duration and involves alterations in respiratory metabolism including accumulation of acetaldehyde and ethanol to high, and possibly toxic, levels during imbibition.