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Query: EC:2.3.1.108 (
TAT
)
2,389
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
We have developed a sensitive assay for leptospira, using the polymerase chain reaction (PCR). On the basis of the published nucleotides sequence of 23S rRNA gene from Leptospira interrogans serovar canicola strain Moulton, primers were chosen to produce an amplified fragment of 123 bp. Primer A: 5'GAT CTA ATT CGC TGT AGC AGG3' and primer B: 5'ACT
TTC
ACC CTC
TAT
GGT CGG3' Eight different svs. of Leptospira interrogans could all be detected by PCR, but the DNAs from L. biflexa. Leptonema bacteria, virus and human could not produce the specific amplified fragment. The assay detected approximately 10 fg of purified leptospiral DNA and 1 microliter serum of experimental animal. Positive results were obtained from simulated positive samples containing a single organism leptospiral DNA. The diagnostic test (proved by "gold standards": Clinical diagnosis; blood culture and MAT) showed that the sensitivity was 92.00%; the specificity 94.35%; the accuracy 92.54%; the positive predictive value 98.17%; the negative predictive value 78.13%; the positive likelihood ratio 16.25; and the negative likelihood ratio 0.0848. The diagnosis of early leptospirosis by using PCR may become a significant addition to diagnostic means.
...
PMID:[Detection of leptospiral DNA in the serum of 175 patients with early leptospirosis by polymerase chain reaction]. 129 12
Mouse immunoglobulin heavy-chain variable region (Ig VH) genes apparently arose from the approximately 600-base-pair-long (approximately 12 tandem repeats of the 48-base-pair-long primordial building block sequence
TTC
-AGC-AGC-CTG-ACT-GGA-
TAT
-GAC-CTG-GAG-TGG-ACT-TAC-TGC-GCA-AGA) that in the original reading frame specified the amino acid sequence Phe-Ser-Ser-Leu-Thr-Gly-Tyr-Asp-Leu-Glu-Trp-Thr-Tyr-Cys-Ala-Arg. The previously identified, shorter prototype building blocks merely represented particular portions of the above primordial sequence. Even today, the direct descendant in toto of this primordial sequence specifies the last one-sixth of each VH coding sequence: the 83rd to 98th amino acid residues. Furthermore, its four truncated derivatives specify the 4th to 14th, 17th to 23rd, 29th to 37th, and 38th to 48th amino acid residues. Accordingly, all three relatively invariant--therefore, conserved--framework regions (FW-1, FW-2, and FW-3) of VHs are specified by recognizable--therefore, conserved--descendants of the primordial sequence.
...
PMID:Identification of the 48-base-long primordial building block sequence of mouse immunoglobulin variable region genes. 680 49
Nature is condemned to play variations of the same theme in evolution, past commitments progressively restricting freedom of choices in evolutionary directions. While each family of genes evolved by the mechanism of gene duplication, this mechanism is extremely inefficient, the usual fate of redundant copies of the ancestral gene being degeneracy. As a result, the euchromatic DNA of higher organisms became a desert in which still-functioning genes are found scattered like oases at an average distance of 35,000 base-pairs of barren stretch between neighbors in the case of mammals. The 20-base-long sequence (AGCTG) (AGCTG) (AGCTG) (GGGTG) can be considered as one of the few ultimate ancestors of all euchromatic DNAs. Long stretches of intergenic spacers are mostly represented by degenerate subfamilies of repeats derived from the above. Certain 30- 50-base-long units of such degenerate subfamilies apparently served as the primordial building block of the ultimate ancestor of each family of genes. For example, the primordial building block of the ancestor for antigen-binding sites (variable regions) of mammalian immunoglobulin heavy chains apparently was
TTC
-AGC-AGC-CTG-ACT-GGA-
TAT
GAC-CTG-GAG-TGG-ACT-TAC-TGC-GCA-AGA, which is the original reading frame specified in the 16-amino-acid-residues-long sequence Phe-Ser-Ser-Leu-Thr-Gly-Tyr-Asp-Leu-Glu-Trp-Thr-Tyr-Cys-Ala-Arg.
...
PMID:Evolution is condemned to rely upon variations of the same theme: the one ancestral sequence for genes and spacers. 682 Jan 35
One hundred and eighty-one families with multiple endocrine neoplasia type 2A (MEN-2A) or familial medullary thyroid carcinoma (FMTC) have been investigated for mutations in the ret protooncogene in Germany. In 8 families with FMTC or MEN-2A, no mutation could be detected in the cysteine-rich domain encoded in exons 10 and 11 of the ret protooncogene. DNA sequencing of additional exons (no. 13-15) revealed rare noncysteine mutations in 3 families (codons 631, 768, and 844). In contrast to these rare events, heterozygous missense mutations in exon 13, codons 790 and 791, were found in 5 families (4 with MTC only; 1 family with MTC and pheochromocytoma) and 11 patients with apparently sporadic tumors. Two different mutations in codon 790 (TTG-->TTT, TTG-->
TTC
; Leu790Phe) and one mutation in codon 791 (
TAT
-->TTT; Tyr791Phe) created a phenylalanine residue. We conclude that codons 790 and 791 of the ret protooncogene represent a new hot spot for FMTC/MEN-2A causing mutations. With the discovery of these considerably common mutations in codons 790 and 791 and the identification of some rare mutations, 100% of the German FMTC/MEN-2A families could be characterized by a mutation in the ret protooncogene.
...
PMID:A new hot spot for mutations in the ret protooncogene causing familial medullary thyroid carcinoma and multiple endocrine neoplasia type 2A. 950 24
We report herein the identification of a new DRB1 allele using sequence-based typing (SBT). This novel allele, HLA-DRB1*11122, was found in an aboriginal individual (SWP71) from the Paiwan tribe in the southern part of Taiwan. This individual was typed by SBT method as having an HLA genotype of HLA-A*24021/24021, HLA-B*4001/4002, HLA-DRB1*11122/15011, HLA-DRB3*0202, and HLA-DRB5*01011. This new allele differs from DRB1*1112 in the polymorphic exon 2 only at codon 34 (CAA-->CAG; both specify glutamine) and from DRB1*1110 in the exon 2 sequence only at codon 32 (CAT-->
TAT
; H32T). The most likely candidate allele which is found in the aboriginal populations of Taiwan and which may mutate into this new allele is DRB1*11011. DRB1*11122 allele differs from DRB1*11011 allele in the polymorphic exon 2 at both codon 34 (CAA-->CAG) and codon 37 (TAC-->
TTC
; T37F). This novel HLA-DRB1*11122 allele was also found in another aboriginal individual (SWP90) from the same Paiwan tribe. This SWP90 individual was typed by SBT method as having an HLA genotype of HLA-A*24021/24021, HLA-B*4002/5502, HLA-DRB1*11122/1201, and HLA-DRB3*01011/0202. However, the original DRB1*1201 sequence from HERLUFF was found to be erroneously reported and the corrected sequence from SWP90 is now presented herein.
...
PMID:New DR5 sequences: a novel DRB1*11122 allele identified in Paiwan tribe members of Taiwan and a corrected sequence for the DRB1*1201 allele. 1170 30
A fragment of Bombyx mori genomic DNA containing one tRNA2Ala gene and one 5S RNA gene has been used to compare the structural features of silkworm 5S RNA and tRNA genes. The nucleotide sequences of both genes and of the primary transcripts produced from them in homologous in vitro transcription systems have been determined. Comparison of the sequences of these two genes with that of another previously analyzed B. mori tRNA2Ala gene reveals common oligonucleotides which may be important transcriptional signals. The oligonucleotides TA(C)
TAT
, AATTTT, and
TTC
are located approximately (+/- 1 nucleotide) 29, 19, and 3 nucleotides, respectively, before the transcription initiation sites of the two tRNA2Ala genes and the one 5S RNA gene we have analyzed. The sequence GGGCGTAG(C)TCAG lies within the coding regions of all three genes. The functional significance of these sequences is suggested by their location within regions required for the transcription of silkworm alanine tRNA genes in vitro.
...
PMID:Silkworm 5S RNA and alanine tRNA genes share highly conserved 5' flanking and coding sequences. 1458 94
Three novel human leukocyte antigen class II alleles (DRB3*0110, DRB1*1140, and DRB1*140102) are described here. The three novel alleles were initially detected as previously unidentified SSO hybridization patterns using CANTYPE((R)) reverse hybridization assay. Sequences were determined by cloning/sequencing. DRB3*0110 allele is identical to DRB3*010101, except for a single nucleotide substitution (CGC-->AGC) changing codon 39 from Arg to Ser. This polymorphism has not, until now, been identified in DRB allele. Thus, this is an unusual mutation as the codon 39 is a fairly conserved region. The new DRB1*1140 is identical to DRB1*1116, except for a single nucleotide substitution at codon 67 from ATC (encoding for isoleucine) to
TTC
(encoding for phenylalanine). This polymorphism is commonly found in DRB1*11 alleles. Compared with DRB1*140101, DRB1*140102 contains a single silent nucleotide substitution (
TAT
-->TAC, both encoding for tyrosine) at codon 78. This polymorphism is commonly found in DRB1*14 alleles. The three new DRB alleles may have been generated by a point mutation event. The DRB3*0110 and DRB1*140102 were identified in Caucasoid individuals. The ethnic origin of the subject carrying the DRB1*1140 allele is Egyptian. The DRB1*140102 was detected in two unrelated individuals; the DRB3*0110 and DRB1*1140 were only identified once, in a total population of 80,000.
...
PMID:Identification of three novel alleles: DRB3*0110, DRB1*1140, and DRB1*140102. 1510 85
An efficient and novel synthesis of bis(tert-butyl)- 1-pyrenylmethyl-silyl group (TBMPS) has been reported having fluorescent properties conferred by the pyrenyl group. This silyl group being base labile is efficiently used for one-pot protection of the 5-OH of the nucleosides. While incorporated terminally at the 5-OH of long sequences viz. AA TGG AGC CAG T and GC
TAT
GTCAGT TCC CCT TGG
TTC
TC, this group is also helpful in subsequent purification by HPLC as well as PAGE. Besides these, a labeled dimer (T*T) and a labeled tetramer (T*TTT) were also synthesized to compare the fluorescence properties of short and long labeled sequences. Fluorescence properties of these sequences were studied in detail to find the suitability of the approach.
...
PMID:One-pot synthesis of TBMPS (bis [tert-butyl)-1 pyrenylmethyl-silyl) chloride as a novel fluorescent silicon-based protecting group for protection of 5'-OH nucleosides and its use as purification handle in oligonucleotide synthesis. 1625 71
Genomic DNA of Blastocystis isolates released into 0.1% Triton X-100 was suitable for amplification and yielded similar results as the genomic DNA extracted with standard kit. The specific B. hominis primers (BH1: GCT
TAT
CTG GTT GAT CCT GCC AGT and BH2: TGA TCC
TTC
CGC AGG
TTC
ACC TAC A) successfully produced the PCR product of about 1,770 bp with all the 7 Blastocystis isolates tested. The restriction fragment length polymorphism (RFLP) patterns yielded by 13 out of 25 restriction endonucleases showed that the 7 isolates could be grouped into 4 subgroups: subgroup-1 consisted of isolate C; subgroup-2 of isolates H4 and H7; subgroup-3 of isolates KP1, Y51 and M12; and subgroup-4 of isolate 27805. The differences between subgroups manifested as clear-cut RFLP patterns. A common band of 230 bp was revealed by Eco R1 in all the Blastocystis isolates tested. The band of about 180 bp was revealed by Alu I, differentiated symptomatic from asymptomatic isolates of this parasite, and might indicate the pathogenicity of this parasite.
...
PMID:Restriction enzyme digestion analysis of PCR-amplified DNA of Blastocystis hominis isolates. 1861 39
Streptococcus pneumoniae is one of the most frequent causative agents of community acquired pneumoniae, meningitis, sinusitis, bronchitis and otitis media both in children and adults. Conventional laboratory methods may sometimes fail to identify S. pneumoniae. The aims of this study were i) to compare the conventional methods and molecular methods which detected pneumococcal surface antigen A (psaA) and autolysin (lytA) genes; ii) to determine the serotype distribution of S. pneumoniae isolated from the respiratory samples. Randomly chosen 62 S. pneumoniae strains isolated from respiratory samples of patients with clinically proven pneumococcal pneumonia (age range: 1-79 years) between years 2000-2006, were included in the study. Classical microbiological analysis for the isolates included Gram staining, optochin sensitivity test performed in 5% CO2 and ambient air and bile solubility test. Capsular serotyping was performed by using latex particles sensitized with mono-specific typing sera (Statens Serum Institut, Denmark). Quellung reaction (Statens Serum Institut, Denmark) was used for serotyping the isolates that gave equivocal results using latex agglutination. Pneumococcal surface antigen A and autolysin genes were detected by in-house polymerase chain reaction (PCR) using psaA1 (5'-CTT TCT GCA ATC ATT CTT G), psaA2 (5'-GCC
TTC
TTT ACC TTG
TTC
TGC), lytAF (5'-ACG CAA TCT AGC AGA TGA AGC) and lytAR (5'-TGT TTG GTT GGT
TAT
TCG TGC) primers. Twenty six different serotypes were detected in 62 S. pneumoniae isolates. The most prevalent capsule serotype was 14 (n= 6), followed by 19A (n= 5). Four isolates could not be typed by the available antisera. All the isolates were optochin sensitive with or without carbondioxide incubation and were bile soluble. All the isolates included in the study have harboured (100%) psaA and lytA genes. No difference was found between the classical and molecular methods for the identification of S. pneumoniae isolates. In conclusion, detection of psaA and/or lytA genes by molecular methods is of value especially in "nonserotypeable strains" when they are performed with conventional methods in clinically proven S. pneumoniae isolates.
...
PMID:[Value of demonstration of pneumococcal surface antigen A and autolysin genes for the identification of Streptococcus pneumoniae clinical isolates]. 1933 75
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