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Fragmentation of rat serum albumin by cyanogen bromide cleavage and the amino acid sequences of four fragments.

Rat serum albumin was cleaved into seven peptides by cyanogen bromide treatment followed by reduction and carboxymethylation. The amino acid composition and the N-terminus of each peptide and the amino acid sequences of four peptides were determined by conventional methods. An alignment of these peptides in the original protein was proposed in the light of the established sequences of bovine and human plasma albumin.

Amino Acid Sequence↗

Cleavage of Trimeresurus flavoviridis phospholipase A2 with cyanogen bromide: sequence of the short peptide fragment and formation of a noncovalently bonded complex from the fragments.

Phospholipase A2 from the venom of Trimeresurus flavoviridis (Habu snake) on treatment with cyanogen bromide is split into less than Glu-Gly-Leu-Trp-Gln-Phe-Asn-Hse greater than, which is derived from the N-terminal moiety and is designated as S-peptide, and the remaining large peptide, which is designated as L-peptide. It should be stressed that the N-terminal residue is pyroglutamyl, unlike other phospholipases A2. The L-peptide alone was about 6% as active as the parent molecule. It occurs in dimeric form, like the parent molecule. When L-peptide was mixed with increasing amounts of S-peptide, the activity increased in a hyperbolic manner, indicating the formation of an ordered complex between L-peptide and S-peptide. The dissociation constant of the complex was 2.1 x 10(-7) M and its specific activity was 2.8 times that of L-peptide.

Amino Acid Sequence↗

Comparison of conventional techniques to differentiate between Taenia solium and Taenia saginata and an improved polymerase chain reaction-restriction fragment length polymorphism assay using a mitochondrial 12S rDNA fragment.

Given the constraints of classical diagnostic methods, i.e., morphological and isoenzymatic studies of proglottids, a polymerase chain reaction test complemented with restriction enzyme analysis has been modified by redesigning one of the primers to reduce nonspecific amplifications experienced when using field samples. The use of these new, highly cestode-specific primers and the restriction enzyme Ddel led to the development of a diagnostic assay that clearly distinguishes between Taenia saginata and T. solium proglottids in field samples. This assay confirms the presence of T. saginata in Ecuador. DNA amplification of some of these taeniids showed different patterns, suggesting the possibility that strain differences exist. These results demonstrate the need for development of useful molecular assays as reliable tools for epidemiological studies on cestodes.

Animals↗

Identification of Nile perch (Lates niloticus), grouper (Epinephelus guaza), and wreck fish (Polyprion americanus) by polymerase chain reaction-restriction fragment length polymorphism of a 12S rRNA gene fragment.

Restriction site analysis of polymerase chain reaction (PCR) products from a conserved region of the 12S rRNA gene has been used for the specific identification of Nile perch (Lates niloticus), grouper (Epinephelus guaza), and wreck fish (Polyprion americanus). Amplification of DNA isolated from muscle samples was carried out using a set of primers flanking a region of 436 bp from the mitochondrial 12S rRNA gene. Digestions of the PCR products with RsaI and Sau96I endonucleases, followed by agarose gel electrophoresis of the digested PCR products, yielded specific profiles that enabled direct identification of each species analyzed.

Animals↗