scholarly journals Development of a multiplex system for determining 11 genetic markers of predisposition to obesity

Author(s):  
G. V. Sergeev ◽  
I. V. Haidukevich ◽  
A. M. Gorkavaya ◽  
M. D. Ameliyanovich ◽  
I. B. Mosse ◽  
...  

A system has been developed to identify 11 genetic markers associated with the risk of obesity: rs10852521, rs11075990, rs1121980, rs1421085, rs1477196, rs17817449, rs3751812, rs7206790, rs8047395, rs9940128 (FTO gene) and rs1137101 (LEPR gene) by minisequencing (SNaPshot analysis). The conditions for carrying out the amplification and minisequencing reactions, as well as the compositions of the reaction mixtures, were optimized so that the analysis was carried out for all 11 markers simultaneously. The resulting system was tested and showed a high degree of reproducibility and sensitivity required for the detection of these polymorphisms.

2019 ◽  
Vol 6 (2) ◽  
pp. 232
Author(s):  
Sutikno Sutikno ◽  
Rudy Priyanto ◽  
Cece Sumantri ◽  
Jakaria Jakaria

ABSTRAK Gen FTO berfungsi sebagai regulasi homeostasis, deposisi lemak dan pengaturan obesitas. Penelitian ini bertujuan untuk mengidentifikasi polimorfisme SNP g.125550A>T di ekson 3 gen FTO pada bangsa sapi potong Indonesia. Sampel darah diperoleh dari 209 ekor sapi, terdiri atas sapi bali (44), madura (20), Pesisir (20), katingan (20), Peranakan ongole (PO) (22), Pasundan (20), Sumba Ongole (SO) (11), brahman (20), simental (15), dan limousin (18). Polimorfisme gen FTO dianalisis menggunakan metode PCR-RFLP (HpyCH4III) dan direct sequencing. Hasil genotiping SNP g.125550A>T adalah polimorfik (genotipe AA, AT, dan TT) pada sapi madura, pesisir, katingan, PO, pasundan, SO, brahman, simental, dan limousin. Frekuensi alel A dan T masing-masing adalah 0,70, 0,68, 0,84, 0,89, 0,70, 0,86, 0,90, 0,73, 0,69 dan 0,30, 0,33, 0,16, 0,11, 0,30, 0,14, 0,10, 0,27, 0,31. Nilai Ho dan He masing-masing adalah 0,60-0,14 dan 0,44-0,18 serta dalam keseimbangan Hardy-Weinberg (P>0.05). Sementara pada sapi bali bersifat monomorfik hanya bergenotipe AA. Hasil sekuensing SNP g.125550A>T ditemukan mutasi tranvesi A menjadi T pada posisi nukleotida  g.125550. Berdasarkan hasil penelitian ini, dapat disimpulkan bahwa SNP 125550A>T gen FTO beragam dan berpotensi dijadikan marka genetik untuk kualitas daging pada bangsa sapi potong Indonesia.Kata Kunci: gen FTO, PCR-RFLP, Sapi, SNP g.125550A>TABSTRACTThe FTO gene functions as regulation of homeostasis, fat deposition and regulation of obesity. This study aimed to identify the polymorphism of SNP g.125550A>T in exon 3 of FTO gene in Indonesian beef cattle. Blood samples were collected from 209 cattle, including bali (44), madura (20), pesisir (20), katingan (20), PO (22), pasundan (20), SO (11), brahman (20), simental (15), and limousin (18). Polymorphism of the FTO gene was analyzed using PCR-RFLP (HpyCH4III) and direct sequencing methods. The results of genotyping SNP g.125550A>T was polymorphic (AA, AT and TT genotypes) in madura, pesisir, katingan, PO, pasundan, SO, brahman, simental, and limousin cattle. The frequency of A and T alleles were 0,70, 0,68, 0,84, 0,89, 0,70, 0,86, 0,90, 0,73, 0,69 and 0,30, 0,33, 0,16, 0,11, 0,30, 0,14, 0,10, 0,27, 0,31 respectively. The values of Ho and He were 0,60-0,14 and 0,44-0,18 respectively and in Hardy-Weinberg equilibrium (P>0,05). While in Bali cattle was monomorphic (AA genotype). Results of sequencing SNP g.125550A>T of the FTO gene found a transverse mutation A to T at the nucleotide position g.125550. As a result of this study, it can be concluded that SNP 125550A>T of the FTO gene was diverse and potentially used as genetic markers for meat quality in Indonesian beef cattle.Keywords: cattle, FTO gene, PCR-RFLP, SNP g.125550A>T.


2010 ◽  
Vol 2 (S1) ◽  
pp. 153-158 ◽  
Author(s):  
Pirmin Nietlisbach ◽  
Alexander Nater ◽  
Maja P. Greminger ◽  
Natasha Arora ◽  
Michael Krützen

1975 ◽  
Vol 23 (3) ◽  
pp. 411 ◽  
Author(s):  
P Clark ◽  
GE Ryan ◽  
AB Czuppon

One quantitative and nine qualitative genetic markers were investigated in some 200 individuals of the family Canidae, including 34 breeds and species. A high degree of homogeneity was observed and only one marker, transferrin, displayed marked variation. The consistent differences were: wolf (one sample), transferrin; jackal (one sample), transferrin and glucosephosphate isomerase; foxes, albumin, transferrin and glucosephsophate isomerase. Two abnormal transferrins were observed in two siblings among six German short-haired pointers. All other markers were homogeneous, usually single protein species or multiple forms dispersed randomly through the group. The results suggest that domestic dogs and dingoes share a common pool of genes and that all canids but the foxes possess many genes in common. There are indications that the jackal and wolf may differ from other canids in some marker systems.


1995 ◽  
Vol 73 (S1) ◽  
pp. 292-301 ◽  
Author(s):  
B. A. McDonald ◽  
R. E. Pettway ◽  
R. S. Chen ◽  
J. M. Boeger ◽  
J. P. Martinez

The DNA-based markers of molecular genetics were combined with the analytical tools of population genetics to learn about the population biology of the wheat pathogen Mycosphaerella graminicola. DNA-based genetic markers, including restriction fragment length polymorphisms in nuclear and mitochondrial DNA, DNA fingerprints, and electrophoretic karyotypes were used in combination to show that the amount and distribution of genetic variation within and among field populations of M. graminicola is similar around the world. Measures of gametic disequilibrium suggested that the sexual stage of reproduction has a more significant impact on the genetic structure of M. graminicola populations than asexual reproduction. A field experiment conducted over a 3-year period showed that populations had a high degree of genetic stability over time. The potential effects of selection were quantified in a cultivar mixture experiment with four wheat cultivars that varied in resistance to M. graminicola. In combination, these experiments demonstrated the utility of selectively neutral genetic markers to elucidate the population genetics of fungi. Key words: genetic diversity, wheat, gene flow, RFLPs, DNA fingerprinting.


Author(s):  
Adrian F. van Dellen

The morphologic pathologist may require information on the ultrastructure of a non-specific lesion seen under the light microscope before he can make a specific determination. Such lesions, when caused by infectious disease agents, may be sparsely distributed in any organ system. Tissue culture systems, too, may only have widely dispersed foci suitable for ultrastructural study. In these situations, when only a few, small foci in large tissue areas are useful for electron microscopy, it is advantageous to employ a methodology which rapidly selects a single tissue focus that is expected to yield beneficial ultrastructural data from amongst the surrounding tissue. This is in essence what "LIFTING" accomplishes. We have developed LIFTING to a high degree of accuracy and repeatability utilizing the Microlift (Fig 1), and have successfully applied it to tissue culture monolayers, histologic paraffin sections, and tissue blocks with large surface areas that had been initially fixed for either light or electron microscopy.


Author(s):  
Cecil E. Hall

The visualization of organic macromolecules such as proteins, nucleic acids, viruses and virus components has reached its high degree of effectiveness owing to refinements and reliability of instruments and to the invention of methods for enhancing the structure of these materials within the electron image. The latter techniques have been most important because what can be seen depends upon the molecular and atomic character of the object as modified which is rarely evident in the pristine material. Structure may thus be displayed by the arts of positive and negative staining, shadow casting, replication and other techniques. Enhancement of contrast, which delineates bounds of isolated macromolecules has been effected progressively over the years as illustrated in Figs. 1, 2, 3 and 4 by these methods. We now look to the future wondering what other visions are waiting to be seen. The instrument designers will need to exact from the arts of fabrication the performance that theory has prescribed as well as methods for phase and interference contrast with explorations of the potentialities of very high and very low voltages. Chemistry must play an increasingly important part in future progress by providing specific stain molecules of high visibility, substrates of vanishing “noise” level and means for preservation of molecular structures that usually exist in a solvated condition.


Author(s):  
P.R. Swann ◽  
A.E. Lloyd

Figure 1 shows the design of a specimen stage used for the in situ observation of phase transformations in the temperature range between ambient and −160°C. The design has the following features a high degree of specimen stability during tilting linear tilt actuation about two orthogonal axes for accurate control of tilt angle read-out high angle tilt range for stereo work and habit plane determination simple, robust construction temperature control of better than ±0.5°C minimum thermal drift and transmission of vibration from the cooling system.


Author(s):  
Willem H.J. Andersen

Electron microscope design, and particularly the design of the imaging system, has reached a high degree of perfection. Present objective lenses perform up to their theoretical limit, while the whole imaging system, consisting of three or four lenses, provides very wide ranges of magnification and diffraction camera length with virtually no distortion of the image. Evolution of the electron microscope in to a routine research tool in which objects of steadily increasing thickness are investigated, has made it necessary for the designer to pay special attention to the chromatic aberrations of the magnification system (as distinct from the chromatic aberration of the objective lens). These chromatic aberrations cause edge un-sharpness of the image due to electrons which have suffered energy losses in the object.There exist two kinds of chromatic aberration of the magnification system; the chromatic change of magnification, characterized by the coefficient Cm, and the chromatic change of rotation given by Cp.


Author(s):  
Robert F. Dunn

Receptor cells of the cristae in the vestibular labyrinth of the bullfrog, Rana catesbiana, show a high degree of morphological organization. Four specialized regions may be distinguished: the apical region, the supranuclear region, the paranuclear region, and the basilar region.The apical region includes a single kinocilium, approximately 40 stereocilia, and many small microvilli all projecting from the apical cell surface into the lumen of the ampulla. A cuticular plate, located at the base of the stereocilia, contains filamentous attachments of the stereocilia, and has the general appearance of a homogeneous aggregation of fine particles (Fig. 1). An accumulation of mitochondria is located within the cytoplasm basal to the cuticular plate.


Author(s):  
E. R. Macagno ◽  
C. Levinthal

The optic ganglion of Daphnia Magna, a small crustacean that reproduces parthenogenetically contains about three hundred neurons: 110 neurons in the Lamina or anterior region and about 190 neurons in the Medulla or posterior region. The ganglion lies in the midplane of the organism and shows a high degree of left-right symmetry in its structures. The Lamina neurons form the first projection of the visual output from 176 retinula cells in the compound eye. In order to answer questions about structural invariance under constant genetic background, we have begun to reconstruct in detail the morphology and synaptic connectivity of various neurons in this ganglion from electron micrographs of serial sections (1). The ganglion is sectioned in a dorso-ventra1 direction so as to minimize the cross-sectional area photographed in each section. This area is about 60 μm x 120 μm, and hence most of the ganglion fit in a single 70 mm micrograph at the lowest magnification (685x) available on our Zeiss EM9-S.


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