Search results for " cloning"

Alternative discourses in science fiction: human cloning in Glory Season (1993) by David Brin

2005

In vivo assembly of chromatin on pBR322 sequences cloned into yeast plasmids

1989

Abstract In order to study the in vivo assembly of chromatin on prokaryotic DNA templates, we have transformed yeast cells with plasmids pAJ50 and pRB58, which contain pBR322 sequences. In both cases nucleosomes are assembled in vivo on pBR322 DNA, although the nucleosomes are not homogeneous in size. To explore whether there is any preference for nucleosome assembly along pBR322 sequences, we have used an indirect end labeling method. The results indicate that most nucleosomes are placed at random on pBR322, although the probability for histone octamers to interact with some short regions is somewhat reduced. These regions coincide with sequences in which the frequency distribution of nucl…

Cloning of cDNAs coding forCandida albicanscell surface proteins

1995

Two cDNA libraries were constructed from mRNAs obtained from yeast cells and germ-tubes of Candida albicans in lambda gt11. Immunoscreening with polyclonal antibodies raised against cell wall components allowed the detection of 29 positive clones. Two of these clones were selected for their specific reactivity with antisera either from yeast (clone 11Y) or germ-tubes (clone 24M). cDNA fragments were isolated by the digestion of lambda DNA with EcoRI. Southern blot analysis with these fragments as probes demonstrated homology with C. albicans DNA, and by Northern analysis two mRNAs transcripts were detected with sizes of approximately 1·5 kb for 11Y and 1·1 kb for 24M. Both transcripts were …

Cloning of an alkane hydroxylase system in a long chain n-alkane- degrader Gordonia sp.

2008

Five Gram-positive GC rich n-alkane degraders were isolated from a long-term accidentally contaminated beach in Sicily and identified as one Nocardia, two Rhodococcus and two Gordonia strains (Quatrini et al., 2008 J. Appl. Microbiol. 104:251-9). All the isolates were able to grow on long and very long chain n-alkanes up to C36. Diverging alkane-hydroxylase encoding genes (alkB) were detected by PCR using degenerated primers in all the strains. Multiple sequences were obtained from the Nocardia strain while only one alkB gene was detected in Rhodococcus and Gordonia. The aim of this work is to genetically characterize the alk cluster in one of the two Gordonia strains called SoCg. Pulsed Fi…

Alcoholismo tipo II, búsqueda de sensaciones y personalidad antisocial : bases neurobiológicas

2000

En el presente trabajo se describen los aspectos fundamentales de la teoría tipológica del alcoholismo de Cloninger, y el apoyo empírico con que cuenta la misma en la investigación actual. A pesar de algunos aspectos críticos que presenta la clasificación de Cloninger, los datos parecen apoyar la existencia de dos tipos básicos de alcoholis¬mo: uno caracterizado por un inicio tardío, menores complicaciones y disfunciones psicológicas, y mejor pronóstico (Tipo I) y otro caracterizado por la presencia de precursores genéticos, inicio más temprano, mayor severidad clínica y peor diagnóstico (Tipo II). Este alcoholismo tipo II presenta una consistente relación con el rasgo de búsqueda de sensac…

Advanced Strategies for Food-Grade Protein Production: A New

2019

Food-grade production of recombinant proteins in Gram-positive bacteria, especially in LAB (i.e., Lactococcus, Lactobacillus, and Streptococcus), is of great interest in the areas of recombinant enzyme production, industrial food fermentation, gene and metabolic engineering, as well as antigen delivery for oral vaccination. Food-grade expression relies on hosts generally considered as safe organisms and on clone selection not dependent on antibiotic markers, which limit the overall DNA manipulation workflow, as it can be carried out only in the expression host and not in E. coli. Moreover, many commercial expression vectors lack useful elements for protein purification. We constructed a “sh…

Molecular cloning and characterization of the complementary DNA coding for the B-chain of murine Clq

1989

AbstractcDNA clones coding for the B-chain of murine Clq were isolated from a mouse macrophage library. The characterized clones include the total coding region plus a leader sequence. High homology was found with human Clq B-chain in the coding region (81%). Northern blot analysis of total RNA from different tissues of Balb/c mice showed one band of approximately 1.2 kb. The highest signal was found in RNA preparations of thioglycolate-activated peritoneal macrophages. The probe also hybridized with mRNA from spleen, thymus and heart. Extremely weak signals were found in liver, kidney, lung and intestine tissues.

Models of Immune Aging

2018

Abstract Biochemical changes, impaired immune responses to new antigens, and inflammation-based disorders are commonly found in aged individuals. Thus, many studies have addressed the immune system of healthy elderly, including centenarians, since a well-preserved immune system appears to be a major factor of longevity. Longitudinal studies in humans are complicated, as most immune changes associated with aging develop slowly. Human models of accelerated immune aging in clinical conditions allow exploring the age-related changes in the human immune system and the mechanisms of accelerated aging in chronic infections and autoimmunity. Even if they do not perfectly mimic immune function and i…

T-cell cloning in human type I diabetes.

1992

Multiplex Ligation-dependent Probe Amplification (MLPA)

2011

The Multiplex Ligation-dependent Probe Amplification (MLPA) is a PCR-based method. The procedure relies on sequence-specific probe hybridization of genomic DNA, followed by multiplex-PCR amplification of the hybridized probe and a semiquantitative analysis of the resulting PCR products. MLPA allows the analysis of around 40 loci in the same reaction, and is a sensitive and relatively fast technique. Only a small amount of DNA is required and results are available within 2 days.The critical factors when performing MLPA analyses from formalin-fixed paraffin-embedded (FFPE) tissues are DNA integrity and purity; for this reason, a suitable DNA extraction method must be chosen.The MLPA protocol …