scholarly journals DNA sequence analysis of I-Ak beta mutants reveals serologically immunodominant region.

1987 ◽  
Vol 166 (2) ◽  
pp. 433-443 ◽  
Author(s):  
B N Beck ◽  
L R Pease ◽  
M P Bell ◽  
J M Buerstedde ◽  
A E Nilson ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Cell Lines ◽  
Variable Region ◽  
Dna Sequence Analysis ◽  
Single Amino Acid ◽  
Single Nucleotide ◽  
Variable Regions ◽  
The Third

We have produced a series of in vitro serologically selected cell lines that express mutant I-Ak molecules. In this report we describe the DNA sequence analysis of the Ak beta gene of four cell lines that express serologically altered Ak beta polypeptides in association with wild-type Ak alpha polypeptides. Each of the major serologic epitopes on the Ak beta polypeptide has been altered in one or more of the four mutants. In addition, the four mutants exhibit a broad spectrum of functional defects when used to stimulate a panel of T hybridomas of various specificities. The DNA sequence analysis revealed that each mutant had sustained a single nucleotide substitution resulting in a single amino acid substitution. All four independent substitutions occurred within or near the third of the four variable regions defined in the beta 1 domain of the A beta polypeptide by allelic comparisons. These data strongly suggest that the third variable region is the major determinant of alloantigenicity on the Ak beta polypeptide.

scholarly journals Cloning and Sequence Analysis of a Highly Polymorphic Cryptosporidium parvum Gene Encoding a 60-Kilodalton Glycoprotein and Characterization of Its 15- and 45-Kilodalton Zoite Surface Antigen Products

2000 ◽  
Vol 68 (7) ◽  
pp. 4117-4134 ◽  
Author(s):  
William B. Strong ◽  
Jiri Gut ◽  
Richard G. Nelson
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Cryptosporidium Parvum ◽  
Diarrheal Disease ◽  
Surface Glycoprotein ◽  
Single Amino Acid ◽  
Gene Encoding ◽  
Single Nucleotide ◽  
Allelic Sequence

ABSTRACT The apicomplexan parasite Cryptosporidium parvum is a major cause of serious diarrheal disease in both humans and animals. No efficacious chemo- or immunotherapies have been identified for cryptosporidiosis, but certain antibodies directed against zoite surface antigens and/or proteins shed by gliding zoites have been shown to neutralize infectivity in vitro and/or to passively protect against, or ameliorate, disease in vivo. We previously used monoclonal antibody 11A5 to identify a 15-kDa surface glycoprotein that was shed behind motile sporozoites and was recognized by several lectins that neutralized parasite infectivity for cultured epithelial cells. Here we report the cloning and sequence analysis of the gene encoding this 11A5 antigen. Surprisingly, the gene encoded a 330-amino-acid, mucin-like glycoprotein that was predicted to contain an N-terminal signal peptide, a homopolymeric tract of serine residues, 36 sites of O-linked glycosylation, and a hydrophobic C-terminal peptide specifying attachment of a glycosylphosphatidylinositol anchor. The single-copy gene lacked introns and was expressed during merogony to produce a 60-kDa precursor which was proteolytically cleaved to 15- and 45-kDa glycoprotein products that both localized to the surface of sporozoites and merozoites. The gp15/45/60 gene displayed a very high degree of sequence diversity among C. parvumisolates, and the numerous single-nucleotide and single-amino-acid polymorphisms defined five to six allelic classes, each characterized by additional intra-allelic sequence variation. The gp15/45/60 single-nucleotide polymorphisms will prove useful for haplotyping and fingerprinting isolates and for establishing meaningful relationships between C. parvum genotype and phenotype.

Identification of clinically relevant yeast species by DNA sequence analysis of the D2 variable region of the 25–28S rRNA gene

Mycoses ◽  
2008 ◽  
Vol 51 (3) ◽  
pp. 209-227 ◽  
Author(s):  
Lorenza Putignani ◽  
Maria G. Paglia ◽  
Eugenio Bordi ◽  
Elena Nebuloso ◽  
Leopoldo P. Pucillo ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Yeast Species ◽  
Variable Region ◽  
Dna Sequence Analysis ◽  
Rrna Gene ◽  
28S Rrna ◽  
28S Rrna Gene

History of Infection With Different Male-Killing Bacteria in the Two-Spot Ladybird Beetle Adalia bipunctata Revealed Through Mitochondrial DNA Sequence Analysis

Genetics ◽  
2002 ◽  
Vol 160 (3) ◽  
pp. 1075-1086 ◽  
Author(s):  
J Hinrich ◽  
G v d Schulenburg ◽  
Gregory D D Hurst ◽  
Dagmar Tetzlaff ◽  
Gwendolen E Booth ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Sequence Analysis ◽  
Dna Sequence ◽  
Dna Sequence Analysis ◽  
Adalia Bipunctata ◽  
Infection Status ◽  
Variable Regions ◽  
Ladybird Beetle ◽  
Male Killing ◽  
History Of

Abstract The two-spot ladybird beetle Adalia bipunctata (Coleoptera: Coccinellidae) is host to four different intracellular maternally inherited bacteria that kill male hosts during embryogenesis: one each of the genus Rickettsia (α-Proteobacteria) and Spiroplasma (Mollicutes) and two distinct strains of Wolbachia (α-Proteobacteria). The history of infection with these male-killers was explored using host mitochondrial DNA, which is linked with the bacteria due to joint maternal inheritance. Two variable regions, 610 bp of cytochrome oxidase subunit I and 563 bp of NADH dehydrogenase subunit 5, were isolated from 52 A. bipunctata with known infection status and different geographic origin from across Eurasia. Two outgroup taxa were also considered. DNA sequence analysis revealed that the distribution of mitochondrial haplotypes is not associated with geography. Rather, it correlates with infection status, confirming linkage disequilibrium between mitochondria and bacteria. The data strongly suggest that the Rickettsia male-killer invaded the host earlier than the other taxa. Further, the male-killing Spiroplasma is indicated to have undergone a recent and extensive spread through host populations. In general, male-killing in A. bipunctata seems to represent a highly dynamic system, which should prove useful in future studies on the evolutionary dynamics of this peculiar type of symbiont-host association.

21 GENERATION OF GGTA1 AND CMAH BIALLELIC KNOCKOUT PORCINE FIBROBLAST CELL LINES BY TRANSCRIPTION ACTIVATOR-LIKE EFFECTOR NUCLEASE AND CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS (CRISPR)/Cas9

2016 ◽  
Vol 28 (2) ◽  
pp. 140
Author(s):  
J.-D. Kang ◽  
S.-M. Ryu ◽  
H.-Y. Zhu ◽  
L. Jin ◽  
W.-X. Li ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Somatic Cell ◽  
Dna Sequence ◽  
Cell Lines ◽  
Gene Knockout ◽  
Fibroblast Cell ◽  
Dna Sequence Analysis ◽  
Transcription Activator ◽  
Catalytic Core ◽  
Fibroblast Cell Lines

The use of porcine organs for pig-to-human transplantation is considered the most promising solution to overcome the growing shortage of human grafts for all transplantation. Unfortunately, human antibodies recognise Gala-1, 3-gal (Gal), and N-glycolylneuraminic acid (Neu5Gc) epitopes on the surface of pig cells, causing hyperacute rejection. Transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 are 2 powerful tools for producing gene knockouts with high efficiency. Here we generate α-1, 3-galactosyltransferase (GGTA1) and cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene knockout porcine fibroblasts cell lines with TALEN and CRISPR/Cas9. Porcine fibroblasts were treated with TALEN and CRISPR/Cas9 designed against the region coding for the catalytic core of GGTA1 and CMAH. A magnetic separation of these cells was used in somatic cell NT. The cloned fetuses and piglets were characterised by T7E1, fluorescent PCR, and DNA sequencing. Nineteen fetuses were harvested using TALEN; 4 of them were GGTA1 gene mutated, and 11 were CMAH gene mutated. The DNA sequence analysis confirmed 5 of 11 are biallelic disruptions of the CMAH gene. For the CRISPR/Cas9, 6 piglets were obtained, and DNA sequence analysis confirmed 1 of 6 is biallelic disruptions of the GGTA1 and 2 of 6 are mono-allelic disruptions of the CMAH gene. We produced several fetuses or piglets via gene editing technology (TALEN and CRISPR/Cas9) and somatic cell NT technology. Furthermore, GGTA1 and CMAH gene knockout porcine fetal fibroblast cell lines derived from the fetus individuals were successfully established. However, expression of α-Gal and Neu5Gc epitopes is needed to be further examined by flow cytometry and confocal microscopy. Our study will provide the possibility of making GGTA1 and CMAH knockout pigs in the future.

scholarly journals DNA inversions between short inverted repeats in Escherichia coli.

Genetics ◽  
1992 ◽  
Vol 132 (2) ◽  
pp. 295-302
Author(s):  
M A Schofield ◽  
R Agbunag ◽  
J H Miller
Keyword(s):  
Escherichia Coli ◽  
Sequence Analysis ◽  
Dna Sequence ◽  
Dna Sequence Analysis ◽  
Inverted Repeats ◽  
Short Sequence ◽  
Site Specific

Abstract Using site-specific mutagenesis in vitro, we have constructed Escherichia coli strains that allow the detection of the inversion of an 800-bp segment in the lac region. The invertible segment is bounded by inverted repeats of either 12 or 23 bp. Inversions occurring at these inverted repeats will restore the Lac+ phenotype. Inversions can be detected at both short homologies at frequencies ranging from 0.5 x 10(-8) to 1 x 10(-7). These events, which have been verified by DNA sequence analysis, are reduced up to 1000-fold in strains deficient for either RecA, RecB or RecC. They are not reduced in strains deficient in the RecF, J pathway. These results show that the RecB,C,D system can mediate rearrangements at short sequence repeats, and probably plays a major role in cellular rearrangements.

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