Pneumococcal conjugate vaccination response in patients after community-acquired pneumonia, differences in patients with S. pneumoniae versus other pathogens.

Objectives: The goal of this study is to investigate the immune response to the 13-valent pneumococcal conjugate vaccine (PCV13) in former pneumococcal CAP patients. We hypothesize that an impaired or suboptimal humoral immune response against (specific) pneumococcal serotypes might explain the vulnerability for pneumococcal disease.

Methods: Hospitalised adult CAP patients who participated in two trials (2004-2006 (n=201) and, 2007-2009 (n=304)) were screened.

Use of gene replacement transformation to elucidate gene function in the qa gene cluster of Neurospora crassa.

Gene replacement by transformation, employing selective genetic recombination techniques, has been used to delete or disrupt the qa-x, qa-y and qa-1S genes of the qa gene cluster of Neurospora crassa. The growth characteristics of the strain carrying the deletion of the qa-y gene support earlier evidence that this gene encodes a quinic acid permease. The strain containing the deletion of the qa-1S gene (delta qa-1S) was examined with respect to quinic acid induction and carbon catabolite repression.

Sequence and characterization of the met-7 gene of Neurospora crassa.

The proteins encoded by the met-7+ and met-3+ genes of Neurospora crassa are required to form a functional cystathionine-gamma-synthase (CGS). The met-7+ gene has been cloned by complementation of a met-7 mutant. The nucleotide sequence of the complementing DNA reveals the presence of a 542-amino acid open reading frame (ORF).

Comparative studies of the quinic acid (qa) cluster in several Neurospora species with special emphasis on the qa-x-qa-2 intergenic region.

The organization of the quinic acid (qa) genes in Neurospora crassa has been compared to that in several other Neurospora species. This gene cluster was found to be highly conserved in all species examined. However, there are numberous restriction fragment length polymorphisms that distinguish the heterothallic and homothallic species.

The Wilhelmine E. Key 1989 invitational lecture. Organization and regulation of the qa (quinic acid) genes in Neurospora crassa and other fungi.

In Neurospora crassa, five structural genes and two regulatory genes control the use of quinic acid as a carbon source. All seven genes are tightly linked to form the qa gene cluster. The entire cluster, which has been cloned and sequenced, occupies a continuous DNA segment of 17.

Bialaphos resistance as a dominant selectable marker in Neurospora crassa.

Conidia of Neurospora crassa are sensitive to the herbicide bialaphos at concentrations of 160 mg/l in Westergaard's or Fries' minimal media. Plasmid pJA4 was constructed by inserting a truncated bar gene from Streptomyces hygroscopicus fused to the his-3 promoter from N. crassa into pUC19.

DNA sequence, organization and regulation of the qa gene cluster of Neurospora crassa.

In Neurospora, five structural and two regulatory genes mediate the initial events in quinate/shikimate metabolism as a carbon source. These genes are clustered in an 18 x 10(3) base-pair region as a contiguous array. The qa genes are induced by quinic acid and are coordinately controlled at the transcriptional level by the positive and negative regulators, qa-1F and qa-1S, respectively.

DNA hybridization analyses of a Gossypium allotetmploid and two closely related diploid species.

The DNAs of two diploid species of Gossypium, G. herbaceum var. africanum (A1 genome) and G.

Expression of qa-1F activator protein: identification of upstream binding sites in the qa gene cluster and localization of the DNA-binding domain.

The qa-1F regulatory gene of Neurospora crassa encodes an activator protein required for quinic acid induction of transcription in the qa gene cluster. This activator protein was expressed in insect cell culture with a baculovirus expression vector. The activator binds to 13 sites in the gene cluster that are characterized by a conserved 16-base-pair sequence of partial dyad symmetry.

Rearrangement mutations on the 5' side of the qa-2 gene of Neurospora implicate two regions of qa-1F activator-protein interaction.

Transcriptional activation of the Neurospora crassa qa genes normally requires the positive regulatory gene, qa-1F+, whose function is controlled by the inducer quinic acid and by the product of the negative regulatory gene, qa-1S+. The properties of qa-1F+ activator have been examined in transcriptional mutations of the qa-2 structural gene, in which activator-independent transcription of qa-2 (qa-2ai mutants) occurs in strains having a qa-1F- gene. Seven qa-2ai mutants with DNA rearrangements in different 5' regions of qa-2 were analyzed in qa-1F+ strains.

Cis-acting and trans-acting regulatory mutations define two types of promoters controlled by the qa-1F gene of Neurospora.

The function of the qa-1F positive regulatory gene of Neurospora has been studied by mapping the initiation sites for transcription of the clustered qa structural genes in wild type, in qa-1F mutants, and in cis-acting activator protein-independent mutants of qa-2 (qa-2ai mutants). Each structural gene under qa-1F control has two to four promoters. The qa-2ai mutations, which include point mutations and small (68-84 bp) duplications 5' to qa-2, allow qa-1F-independent transcription from surrounding qa promoters independently of the orientations and positions (up-stream or downstream) of teh mutations relative to the promoters.

Point mutations and DNA rearrangements 5' to the inducible qa-2 gene of Neurospora allow activator protein-independent transcription.

Expression of the qa-2 gene of Neurospora crassa normally requires a functional activator protein encoded by qa-1F. Twelve transcriptional mutants of the qa-2 gene have been isolated in qa-1F- strains, and these allow partial expression of qa-2 (1-45% of induced wild type) in the absence of functional activator protein. All 12 mutants have been characterized by genomic (Southern) blot hybridization and the DNAs of 5 have been cloned and sequenced.

Chimeric plasmid that replicates autonomously in both Escherichia coli and Neurospora crassa.

A hybrid pBR322 plasmid (designated pDV1001) containing two functional Escherichia coli antibiotic resistance genes (kanr and camr) and a qa-2+ gene from Neurospora crassa transforms N. crassa qa-2- mutants to qa-2+ with a frequency of ca. 5 X 10(-5) per regenerated spheroplast (ca.

Direct identification of sickle cell anemia by blot hybridization.

Several reports have been published on the use of polymorphisms found in the human hemoglobin genes as a means for prenatal diagnosis of sickle cell anemia. The disadvantages of this approach reside in its limited application and the need for family analysis. Here we report that, by use of restriction endonuclease Dde I and diazobenzyloxymethyl-paper transfer procedures, a direct analysis can be made.