scholarly journals Novel primer targeting the mitochondrial NADH dehydrogenase subunit 4 (ND4) and NADH dehydrogenase subunit 5 (ND5) for detection of porcine (Sus scrofa) DNA fragments in food products for halal authentication

2021 ◽  
Vol 924 (1) ◽  
pp. 012004
Author(s):  
J Kusnadi ◽  
P Mahardita ◽  
K U Al-Awwaly ◽  
E L Arumingtyas

Abstract In this research, a pair primer targeting NADH dehydrogenase subunit 4 (ND4) gene of porcine (Sus scrofa) mt-DNA was designed and tested for its specificity. The ND4 primer was compared for its robustness against an established primer pair designed to amplify the NADH dehydrogenase subunit 5 (ND5) gene. The samples include non-halal meat animals (dog, porcine, and rat) and halal meat animals (chicken, cow, and horse). DNA from raw meat was isolated by modified Chloroform Isoamyl-Alcohol method and then was analyzed quantitatively with NanoDrop™ spectrophotometer followed by amplification using PCR technique. The amplification results proved that the two pairs of primers were specific, resulting in amplification of 120 bp DNA target for ND4 and 227 bp for ND5. It can be concluded that the two primers can differentiate between halal and non-halal animals. However, to determine the sensitivity of each primer, further research is needed.

Genetics ◽  
1998 ◽  
Vol 148 (1) ◽  
pp. 409-421 ◽  
Author(s):  
Cheryl A Wise ◽  
Michaela Sraml ◽  
Simon Easteal

Abstract To test whether patterns of mitochondrial DNA (mtDNA) variation are consistent with a neutral model of molecular evolution, nucleotide sequences were determined for the 1041 bp of the NADH dehydrogenase subunit 2 (ND2) gene in 20 geographically diverse humans and 20 common chimpanzees. Contingency tests of neutrality were performed using four mutational categories for the ND2 molecule: synonymous and nonsynonymous mutations in the transmembrane regions, and synonymous and nonsynonymous mutations in the surface regions. The following three topological mutational categories were also used: intraspecific tips, intraspecific interiors, and interspecific fixed differences. The analyses reveal a significantly greater number of nonsynonymous polymorphisms within human transmembrane regions than expected based on interspecific comparisons, and they are inconsistent with a neutral equilibrium model. This pattern of excess nonsynonymous polymorphism is not seen within chimpanzees. Statistical tests of neutrality, such as Tajima's D test, and the D and F tests proposed by Fu and Li, indicate an excess of low frequency polymorphisms in the human data, but not in the chimpanzee data. This is consistent with recent directional selection, a population bottleneck or background selection of slightly deleterious mutations in human mtDNA samples. The analyses further support the idea that mitochondrial genome evolution is governed by selective forces that have the potential to affect its use as a “neutral” marker in evolutionary and population genetic studies.


Parasitology ◽  
2004 ◽  
Vol 130 (2) ◽  
pp. 239-246 ◽  
Author(s):  
J.-Y. CHUNG ◽  
W.-G. KHO ◽  
S.-Y. HWANG ◽  
E.-Y. JE ◽  
Y.-T. CHUNG ◽  
...  

Acephalic cysticercus (Ac), a rarely developed multilobulated and nonencysted form of larval Taenia, causes hydrocephalus or adhesive arachnoiditis in the ventricles and subarachnoidal space that often lead to fatal outcome in affected patients. Ac has been proposed to originate from T. solium on the basis of morphological features, while no molecular data supporting the presumption have been available. In the present study, we investigated the immunological properties as well as molecular characteristics of Ac that was obtained surgically from 6 patients. Immunoblotting of the cyst fluid from Ac samples demonstrated the constitutive expression of a T. solium metacestode (TsM) 10 kDa protein. Specific antibodies against the truncated 10 kDa protein, which appears to be species specific for TsM cysticercosis, were detected in both serum and cerebrospinal fluid samples of Ac patients. Nucleotide sequences of mitochondrial cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 1 (ND1) genes of Ac were almost identical to those of T. solium but differed substantially from those of the other Taenia species. In phylogenetic analysis, Ac clustered with T. solium in a well-supported clade. Our results strongly suggest that Ac may have originated from T. solium.


FEBS Letters ◽  
2005 ◽  
Vol 579 (16) ◽  
pp. 3449-3460 ◽  
Author(s):  
Madhubanti Sarkar ◽  
Soumita Das ◽  
Arunava Bandyopadhaya ◽  
Kunal Ray ◽  
Keya Chaudhuri

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Mervat M. Abo-Elkhier ◽  
Marwa A. Abd Elwahaab ◽  
Moheb I. Abo El Maaty

The comparison of protein sequences according to similarity is a fundamental aspect of today’s biomedical research. With the developments of sequencing technologies, a large number of protein sequences increase exponentially in the public databases. Famous sequences’ comparison methods are alignment based. They generally give excellent results when the sequences under study are closely related and they are time consuming. Herein, a new alignment-free method is introduced. Our technique depends on a new graphical representation and descriptor. The graphical representation of protein sequence is a simple way to visualize protein sequences. The descriptor compresses the primary sequence into a single vector composed of only two values. Our approach gives good results with both short and long sequences within a little computation time. It is applied on nine beta globin, nine ND5 (NADH dehydrogenase subunit 5), and 24 spike protein sequences. Correlation and significance analyses are also introduced to compare our similarity/dissimilarity results with others’ approaches, results, and sequence homology.


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