The underwater light field in the Bellingshausen and Amundsen Seas (Antarctica)

The underwater light field in the Bellingshausen andAdmundsen Seas was characterised using data collectedduring the R/V Polarstern cruise ANT XI/3, from12.1.94 to 27.3.94. The euphotic zone varied from 24to 100 m depth. Spectral diffuse vertical attenuationcoefficients (K _d ())were determined for 12narrow wavebands as well as for photosyntheticallyavailable radiation (PAR, 400–700 nm): K _d (490)ranged from 0.03 to 0.26 m¹; K _d (550) from0.04 to 0.17 m¹; K _d (683) from 0.04 to0.17 m¹; and K _d (PAR) varied from 0.02 to0.25 m¹. K _d () for wavelengths centred at412 nm, 443 nm, 465 nm, 490 nm, 510 nm, 520 nm and550 nm were significantly correlated with chlorophyllconcentration (ranging from 0.1 to 6 mg m³). Thevertical attenuation coefficients for 340 nm and380 nm ranged from 0.10 to 0.69 m¹ and from 0.05to 0.34 m¹, respectively, and were also highlycorrelated with chlorophyll concentrations. These K _d values indicate that the 1% penetration depthmay reach maxima of 46 m and 92 m for 340 nm and380 nm, respectively. The spectral radiancereflectances (Rr()) for 443 nm, 510 nm and 550 nmwere less than 0.01 sr¹. Rr() for 665 nm and683 nm increased with depth up to 0.2 sr¹ because ofchlorophyll fluorescence. Using a model that predicts downwardirradiances by taking into account the attenuation bywater and absorption by chlorophyll, we show thatchlorophyll fluorescence has a significant influenceon the red downward irradiance (E _d (633, 665, 683))in deeper layers. The ability of the phytoplanktonpopulation to influence the light environment byautofluorescence and absorption processes depends onthe light conditions and on the photoacclimation ofthe cells, represented by the in vivo crosssection absorption coefficient of chlorophyll (a*). Theobtained mean chlorophyll-specific light attenuationcoefficients of phytoplankton in situ (k _d ) are higherthan the in vivo absorption coefficient of chlorophyll,more than to be excepted from the scattering. a*(), m² mg chl¹, decreased due topackaging effect with increasing chlorophyllconcentrations.     

Lambda phage mutants insensitive to temperature-sensitive repressor

Summary Weak-virulent mutants of temperate coli-phage were isolated which can grow on the CIts lysogen producing a temperature-sensitive repressor but which cannot grow on the wild type lysogen producing a normal repressor.Genetic analysis on the mutants shows that their weak-virulence is attributable to two mutations, one (virL) in the region between sus N₂₁₃ and c ₄₇ and the other (virR) in the region between c ₁ and sus O₈. Both mutations are located within the region of non-homology between and imm ⁴³⁴ phages.True virulent mutants which can grow on the wild type lysogen can be obtained easily from the weak-virulent mutant by an additional mutation, virC in a region very close to virR. The virulent mutants obtained are similar to the classical vir mutant (Jacob and Wollman, 1954). The virL and virR mutations are probably operator mutations which render the genome insensitive to the repressor.This work was reported at the XII th International Congress of Genetics, held in Tokyo, on August 23, 1968.     

Rearrangements between the operators in the bacteriophage lambda

Summary The left operator mutant v2s develops poorly during infection as a result of constitutive expression of the left operon. A revertant of v2s, designated iri, was found to contain an inversion of the cI region with the inversion endpoints to be within the lambda operators o _L and o _R. Formation of the inversion is facilitated by a translocation of right operator o _R ^c mutant sequence to the left operator in v2s. The inversion in iri positions wild-type o _R sequence at o _L returning control of the left operon to repression by the lambda cro repressor.     

The spectral input systems of hymenopteran insects and their receptor-based colour vision

Summary Spectral sensitivity functions S() of single photoreceptor cells in 43 different hymenopteran species were measured intracellularly with the fast spectral scan method. The distribution of maximal sensitivity values (_max) shows 3 major peaks at 340 nm, 430 nm and 535 nm and a small peak at 600 nm. Predictions about the colour vision systems of the different hymenopteran species are derived from the spectral sensitivities by application of a receptor model of colour vision and a model of two colour opponent channels. Most of the species have a trichromatic colour vision system. Although the S() functions are quite similar, the predicted colour discriminability curves differ in their relative height of best discriminability in the UV-blue or bluegreen area of the spectrum, indicating that relatively small differences in the S() functions may have considerable effects on colour discriminability. Four of the hymenopteran insects tested contain an additional R-receptor with maximal sensitivity around 600 nm. The R-receptor of the solitary bee Callonychium petuniae is based on a pigment (P596) with a long _max, whereas in the sawfly Tenthredo campestris the G-receptor appears to act as filter to a pigment (P570), shifting its _max value to a longer wavelength and narrowing its bandwidth. Evolutionary and life history constraints (e.g. phylogenetic relatedness, social or solitary life, general or specialized feeding behaviour) appear to have no effect on the S() functions. The only effect is found in UV receptors, for which _max values at longer wavelengths are found in bees flying predominantly within the forest.     

Inactivation of DNA-binding activity of repressor in extracts of lambda-lysogen treated with mitomycin C

Summary Crude extracts from -lysogens treated with mitomycin C were prepared, and immunity repressor levels in the extracts were assayed by the binding activity specific to DNA immunity region. It has been shown that while the repressor levels in the extracts from C600(+) are reduced after mitomycin C treatment, the levels in C600recA(+) and C600 C72(+) which have defects in lifting the immunity are not affected by the treatment. The repressor levels in the extracts prepared from C600 T44(+) after temperature shift up, whose prophage is inducible at high temperature, are also reduced. From the study of chloramphenicol effect, it was indicated that de novo protein synthesis is required for the inactivation of the repressor in C600(+) by mitomycin C, but not in T44(+) by high temperature.     

Structure and function of the repressor of bacteriophage lambda

Summary By mutagenizing a cIts (cI857) lysogen, a mutant has been isolated with a wild-type phenotype. This mutant phage lysogenizes with low efficiency and produces a low burst. Though the initial rates of repressor synthesis in Escherichia coli after infection with wild-type and mutant are the same, the maximum level of repressor that is synthesized in the latter case is only about 30% of that synthesized in the former. Virulent plates on the lysogen of mutant with slightly less efficiency producing very tiny plaques. Operator-binding studies made in vitro with purified mutant and wild-type repressors show that the binding curve of the former repressor is a rectangular hyperbola while that of the latter is sigmoid. The half-lives of the complexes of mutant and wild-type repressors with right operator are 133 and 27 min, respectively. All these results suggest that the mutant repressor possibly has a higher affinity for the operators. This mutant has been named cIha (ha=high affinity).     

Cloning of the replication gene P of bacteriophage lambda: Effects of increased P-protein synthesis on cellular and phage DNA replication

Summary A restriction fragment of DNA carrying the P gene was cloned in the high copy number plasmid RSF2124. Cells harbouring this new plasmid RSF2124/E complement Pam80 phage. A lac promoter-operator region (lacP), produced by EcoRI digestion of plasmid pKB252, was inserted into RSF2124/glE such that induction of the lac promoter by IPTG or lactose leads to increased production of the P gene product. A high amount of P protein in E. coli cells results in a slow inhibition of bacterial DNA synthesis, suggesting that the initiation reaction is blocked by P protein. Synthesis of DNA proceeds normally under these conditions.Nonsuppressing groPA15 mutant bacteria which are unable to support the replication of wild-type (wt), acquire the ability to replicate Pam80 phage but not wt when they are transformed with a plasmid carrying the P gene. When harbouring a plasmid containing the mutant Pamber 80 gene, groPA15 mutants are able to support the replication of wt phage when infected at a high multiplicity. Pam80 phage does not multiply in these cells.     

Fertility functions of resistance factors

Summary The isolation of transducing phages carrying the tolPAB cluster is described. These genes map between gltA and gal in Escherichia coli, and thus are relatively close to att. To isolate these transducing phages, it was necessary to use a strain deleted of most of the intervening genes (nadA to chlD) between tolPAB and att. Using a lysogen of such a deletion strain, several defective dtol phages were isolated that carry different amounts of the tolPAB cluster.All of these dtolPAB phages were defective in both lysogenization and vegetative growth, and in this respect were similar to dgal transducing phages.The usefulness of such specialized transducing phages in studying the cell surface is discussed.Research Fellow of the National Cancer Institute of Canada.     

Analysis of lambda receptor and beta-lactamase synthesis and export using cloned genes in a minicell system

Summary We have cloned lamB, the gene for receptor (an outer membrane protein), on a small plasmid which also carries the gene for -lactamase (a periplasmic protein). We have identified a promoter in the region of malK, the gene immediately preceding lamB, which is active in minicells but relatively inactive in vitro. Using a minicell system, we have found that both receptor and -lactamase are made as full length precursors which are subsequently processed. We also show that the receptor precursor can be exported to the outer membrane before it is processed. Mature -lactamase is found only in the periplasm, suggesting that processing may be a requirement for export to the periplasm.     

On the mode of antirepressor action in anti-immune cells

Summary The mode of antirepressor action in anti-immune cells was analysed in respect to its two main features, low lysogenization and high cell killing. By means of complementation experiments between and i434 in anti-immune cells, it was shown that the antirepressor no longer channels phages towards lysis in such cells if the genes which are needed for lysogenization are provided in trans by the heteroimmune phage i434. Since complementation could be demonstrated, it was possible to exclude that direct action of the antirepressor over repressor production is responsible for the feature under analysis. It was also shown that both int- and cII-product are required to improve lysogenization and to prevent high levels of killing.Recipient of an EMBO predoctoral fellowship.     

Analysis of a temperature sensitive mutation in gene cII of bacteriophage lambda

Summary The mutation cIIts612 was found to map outside the immunity region of phage imm21 hybrid. As expected of a cII mutation, cIIts612 is unable to stimulate either cI repressor or Int synthesis during the establishment of lysogeny. These results indicate that part of the cII gene of is homologous to that of imm21 phage.     

Availability of oncogene activated production system for mass production of light chain of human antibody in CHO cells

We previously established a ras-oncogene amplified Chinesehamster ovary (CHO) cell line, named ras clone I, as anuniversal host cell line for oncogene activated production(OAP) system to mass-produce recombinant protein by activationof the cytomegalovirus immediate early (CMV) promoter with ras protein. The light chain(C5) of human monoclonal antibody HB4C5 is expected tobe potentially useful for lung cancer targeting. We generated aC5 hyper-producing cell line by transfecting ras cloneI with the C5 gene expression plasmid regulated by theCMV promoter, of which productivity was 5.3 times greater thanthe hyper productive CHO cell line generated by using conventional CHO cells. Introduction of the adenovirus E1A geneinto the hyper-producing cell line derived from ras clone I resulted in further 9.5 times enhancement of the productivity,suggesting the synergistic effect of E1A and ras oncogenes on the recombinant protein production driven by the CMV promoter. In addition, intracellular accumulation of C5 andupregulation of BiP was found in hyper-producing cell lineswhich were introduced E1A and ras oncogene. This resultsuggests that excessive intracellular accumulation ofC5 protein, which might be caused by that the amount of produced C5 in ER is beyond the ability of CHO cells to secrete, might signal the BiP promoter. Our data imply that ras clone I is available as a general host cell for establishing the recombinant protein hyper-producing CHOcells by the OAP system, and suggest that further mass production of recombinant proteins in the OAP system can be possible by clarifying the accurate role of upregulated BiP protein.     

Two insertions in the galactose operon having different sizes but homologous DNA sequences

Summary DNA from dg phages carrying the strong-polar insertions N 102 and N 116 was transcribed into RNA in vitro. The RNA was exhaustively hybridized to dg-DNA carrying the wildtype galactose operon. The remaining RNA contained a fraction which specifically hybridized to DNA carrying the insertion.The insertion-specific RNA hybridizes equally well to both insertions, regardless from which insetion the RNA was transcribed. The amount of insertion-specific RNA transcribed from the larger insertion N 102 is 2–3 times larger than the amount transcribed from insertion N 116.Herrn Prof. Dr. Hans Netter zum 70. Geburtstag gewidmet.     

Construction and some properties of packageable plasmid F

Summary A derivative of plasmid F which is packageable in phage coat was constructed using techniques of in vitro recombination. This plasmid is composed of three DNA fragments generated by restriction enzyme EcoRI: a miniF fragment (fragment f5 of F'lac) which is able to replicate autonomously, a DNA fragment from Staphylococcus Plasmid that carries the -lactamase gene, and a portion of guaA (B) transducing phage DNA carrying cohesive ends (cos site) along with almost all the late genes but devoid of all those genes and sites that are needed for replication, regulation, and recombination. The hybrid plasmid has a molecular weight of 2.7×10⁷ daltons, about 84% size of phage genome, and can be packaged in coat when helper phage replicates in the plasmid-carrier cell. The package plasmid and the helper phage particles are separated by CsCl density gradient centrifugation. The replication characteristics of the recombinant plasmid are all those of F including the copy number, incompatibility, and curing with acidine orange. The packaged plasmid is injected into an F^- cell and establishes a plasmid state with normal efficiency. In F⁺ or Hfr cells, the resident F factor hinders this process.     

Structure and molecular organization of the photosynthetic accessory pigments of cyanobacteria and red algae

Summary Cyanobacteria (blue-green algae) and Rhodophyta (red algae) contain high concentrations of photosynthetic accessory pigments (phycobiliproteins) which trap light energy in the region between 400 and 650 nm. The electronic excitation energy is then transferred along a chain of these pigments to the reaction center chlorophyll of Photosystem II by a radiationless induced resonance process.Unlike the protein-chlorophyll complexes in the photosynthetic lamellae, the phycobiliproteins are readily soluble in aqueous solution, can be isolated in a variety of assembly forms, and crystallize readily. These properties facilitate the study of the structure of these proteins by chemical, physical, and immunological methods, as well as by X-ray diffraction and electron microscopy.The brilliantly colored phycobiliproteins are a homologous family of conjugated proteins of differing spectroscopic properties. The basic structural unit in these proteins is a monomer of 30,000–40,000 daltons made up of two dissimilar polypeptide chains, and . Each subunit carries covalently linked tetrapyrrole prosthetic groups related to the bile pigment biliverdin.The distinctive spectroscopic properties of each phycobiliprotein are a consequence of the chemical structure of the bile pigment it carries, and of the influence of the conformation and aggregation state of the protein on the spectra of these prosthetic groups. In vivo, the phycobiliproteins are organized into particles, phycobilisomes, attached in a regular array to the outer surface of the photosynthetic lamellae. Studies on phycobilisomes, and on intact cells, indicate the following pathway of energy transfer.Phycoerythrin Phycocyanin (_max 560 nm) (_max 620 nm) Allophycocyanin Allophycocyanin B (_max 650 nm)(_max 671 nm) Chlorophyll a (_max 680 nm)The amounts of the various phycobiliproteins in the cell are influenced by the intensity and energy distribution of the incident radiation. The phenomena of intensity adaptation and complementary chromatic adaptation yield insights into the structure of phycobilisomes and the molecular basis of the plasticity of the structure of this light-harvesting system.Invited article.     

Lambda encodes an outer membrane protein: the lom gene.

Summary infected minicells synthesize a polypeptide (M _r=20,500) which is incorporated almost exclusively into the outer membrane of the minicell envelope. The gene (lom=lambda outer membrane) encoding this polypeptide has been mapped in the nonessential region of the genome between coordinates 39.4% and 40.7% of .     

An improved selection method for λlacZ − phages based on galactose sensitivity

The determination of thelacZ mutant frequency in gt10lacZ phage vectors isolated from the transgenic mouse strain 40.6 (MutaMouse), requires the screening of large numbers of phages on -galactosidase activity. Existing methods rely on distinguishing a few white plaques on X-gal containing plates amongst a multide of blue ones which is both time-consuming and expensive. The new screening method described here employs the galactose sensitiveEscherichia coli C lacZ recA galE strain into which a multicopy plasmid has been introduced, which results in over-expression of thegalK andgalT genes. In the presence of phenyl--d-galactopyranoside, a substrate for -galactosidase, this leads to the suppression of lacZ ⁺ phage propagation without affecting the ability of lacZ ^– phages to form plaques. With this method it is possible to screen 1.5×10⁶ phages on a single 9-cm Petri dish. Furthermore, the need for blue/white screening has been eliminated.     

Human λ light chain locus: Organization and DNA sequences of three genomicJ regions

Evidence for the genomic organization of human lambda light chain joining (J) region gene segments is presented. A mouse J probe was used in Southern hybridizations to localize joining region sequences in a cosmid clone containing the genomic cluster of six human lambda constant (C) region gene segments. The results of these hybridizations suggest the presence of at least one J gene segment upstream from each constant region gene segment. The DNA sequences indicate that the human JI, J2, and J3 gene segments have consensus nonamer and heptamer sequences, proposed to be involved in V-J joining, are capable of encoding the known amino acid sequences for the respective J peptides, and have a sequence which could give functional RNA splice site at the end of their coding regions. Our data show that a single functional J is located 1.3 or 1.6 kb upstream of each of the C gene segments known to encode the Mcg, Kern^– Oz^–, and Kern^–Oz⁺ isotypes. Therefore, the gene organization of this region of the human lambda locus is J1 CI -J2C2-J3C3. The DNA sequences ofJ 1,J 2, andJ 3 presented in this paper establish that a singleJ gene segment precedes each expressed C gene segment, and support a model for the evolution of the human JC clusters where JICI andJ2C2-J3C3. arose from different ancestral JC units.     

Mapping of gal minus-mutants by transducing lambda dg carrying deletions of the gal-region

Summary To map numerousgal ^--mutants ofE. coli, advantage is taken of the fact that transducing dg's can carry different amounts of bacterial DNA of the host from which they originated (Adler andTempleton, 1963).A method is described with which a large number of transducing dg can be easily isolated, differing from each other with respect to the amount of bacterial DNA of thegal-region. By observing whethergal ⁺-colonies can arise as the result of recombination betweengal ^--mutants and dg's carrying deletions in thegal-region, so far 104 kinaseless mutants and 96 transferaseless mutants could be ordered into 26 groups. The mapping-tests were done by spotting the mutants with 52 HFT-lysates of dg's lacking more or less of the kinase- or the kinase- and transferase gene.