A cattle genetic linkage map was constructed which covers more than 95 percent of the bovine genome at medium density. Seven hundred and forty six DNA polymorphisms were genotyped in cattle families which comprise 347 individuals in full sibling pedigrees. Seven hundred and three of the loci are linked to at least one other locus. All linkage groups are assigned to chromosomes, and all are orientated with regards to the centromere. There is little overall difference in the lengths of the bull and cow linkage maps although there are individual differences between maps of chromosomes. One hundred and sixty polymorphisms are in or near genes, and the resultant genome-wide comparative analyses indicate that while there is greater conservation of synteny between cattle and humans compared with mice, the conservation of gene order between cattle and humans is much less than would be expected from the conservation of synteny. This map provides a basis for high-resolution mapping of the bovine genome with physical resources such as Yeast and Bacterial Artificial Chromosomes as well as providing the underpinning for the interpolation of information from the Human Genome Project.

BackgroundHelicobacter pylori is a major human pathogenic bacterium in gastric mucosa. Although the association between gastric cancer and H. pylori has been well-established, the molecular mechanisms underlying H. pylori-induced carcinogenesis are still under investigation. MicroRNAs (miRNAs) are small noncoding RNAs that modulate gene expression at the posttranscriptional level. Recently, studies have revealed that miRNAs are involved in immune response and host cell response to bacteria. Also, microRNA-375 (miR-375) is a key regulator of epithelial properties that are necessary for securing epithelium-immune system cross-talk. It has been recently reported that miR-375 acts as an inhibitor of H. pylori-induced gastric carcinogenesis. There are few reports on miRNA-mediated targeting long noncoding RNAs (lncRNAs).ObjectivesThis study aimed to examine the possible effect of miR-375 as an inhibitor of H. pylori-induced carcinogenesis on the expression of lncRNA SOX2 overlapping transcript (SOX2OT) and SOX2, a master regulator of pluripotency of cancer stem cells.Materials and MethodsIn a model cell line, NT-2 was transfected with the constructed expression vector pEGFP-C1 contained miR-375. The RNA isolations and cDNA synthesis were performed after 48 hours of transformation. Expression of miR-375 and SOX2OT and SOX2 were quantified using real-time polymerase chain reaction and compared with control cells transfected with pEGFP-C1-Mock clone. Cell cycle modification was also compared after transfections using the flow cytometry analysis.ResultsFollowing ectopic expression of miR-375, SOX2OT and SOX2 expression analysis revealed a significant decrease in their expression level (P < 0.05) in NT-2 cells compared to the control. Cell cycle analysis following ectopic expression of miR-375 in the NT-2 cells using propidium iodine staining revealed significant extension in sub-G1 cell cycle.ConclusionsThis is the first report to show down-regulation of SOX2OT and SOX2 following induced expression of miR-375. This finding may suggest expression regulation potential between different classes of ncRNAs, for example between miR-375 and SOX2OT. This data not only extends our understanding of possible ncRNA interactions in cancers but also may open novel investigation lines towards elucidation of molecular mechanisms controlling H. pylori inflammation and carcinogenesis.

Some studies showed analysis of fetal DNA in maternal plasma had been introduced as a new method for non-invasive prenatal diagnosis. Fetal sexing is possible at 8th week of pregnancy, using maternal blood sample testing. The aim was providing a rapid, reliable and non-invasive method for sexing of bovine fetuses. Maternal blood samples were collected from 38 pregnant cows during the 8 th -38 th w of gestation. Plasma was obtained by centrifugation and DNA was extracted by phenol-chloroform method from 350 μL maternal plasma. Two primer pairs for bovine amelogenin, Y-encoded, and testis-specific protein gene were used to amplification three fragments; 260 bp (Y-encoded, testis-specific protein gene), 341 and 467 bp (Y and X chromosome amelogenin gene). The polymerase chain reaction has been optimized for fragments amplification. The 467 bp fragment was detected in all samples. The 341 and 260 bp fragments were detected in 24 of 38 plasma samples of cows with male fetuses. The sensitivity and specificity of test was 100% with no false negative and positive results. The results showed that phenol-chloroform method was a simple and sensational to isolation fetal DNA in maternal plasma. The polymerase chain reaction is a favorable noninvasive method for bovine fetal sexing.

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