Thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) was performed to amplify the uncharacterized regions adjacent to the nusG-rplKAJL-rpoB gene cluster of citrus greening organism (GO) isolates from different locations in Japan and Indonesia. Conventional PCR was used to amplify the internal nusG-rplKAJL-rpoB gene cluster of these isolates, and the complete sequence of this 6.1-kb fragment was determined. Comparisons with other bacterial sequences showed that the fragment is the tufB-secE-nusG-rplKAJL-rpoB gene cluster. The organization of this gene cluster is similar to that of the homologous cluster found in Escherichia coli. Except for three nucleotide changes, the sequence was identical among Japanese and Indonesian isolates. A loop-mediated isothermal amplification (LAMP) assay based on the conserved sequence of the nusG-rplKAJL-rpoB gene cluster was developed for the detection of the GO. The LAMP product was rapidly detected on nylon membranes by staining with AzurB. LAMP could detect as low as about 300 copies of the nusG-rplKAJL-rpoB fragment of the Japanese and Indonesian isolates of GO. The LAMP-based detection method, which does not depend upon a thermal cycler and electrophoresis apparatus, will be useful for under-equipped laboratories, including those found in extension centers and quarantine offices.
It appears that KK-A(y)/Ta mice, especially in terms of histopathological findings, are a suitable animal model for the early stage of type 2 diabetic nephropathy.
The present study was conducted to elucidate the effect of dietary lysine levels on the intramuscular fat (IMF) content in the Longissimus dorsi (L. dorsi) muscles of finishing gilts. Eleven gilts in total from two litters of pigs aged 110 days were used. The average initial bodyweight of the pigs was 61.7 kg. Six pigs were assigned to the low lysine (LL) diet group (lysine content: 0.43 or 0.40%) and five pigs were assigned to the control group (lysine content: 0.65 or 0.68%). The diets were iso-energetic and iso-protein, and contained all essential amino acids (apart from lysine) in the recommended amounts. The pigs were fed these diets until their live weights reached 110 kg. Live weight gain and feed efficiency tended to be lower in the LL group ( P = 0.118 and P = 0.052, respectively). Pigs from the LL group took 5 days longer to reach 110 kg ( P < 0.01). The IMF content in the L. dorsi of the LL group was twice as high as that of the control group (6.7 vs 3.5%; P < 0.01). The percentage of oleic acid in the L. dorsi of the LL group tended to be higher than that of the control group ( P = 0.052), whereas the percentage of linoleic acid and the total percentage of polyunsaturated fatty acids in the L. dorsi were lower ( P < 0.05) in the LL group. Free L-carnitine content in the L. dorsi was lower ( P < 0.05) in the LL group. The average abundance of peroxisome proliferator-activated receptor gamma mRNA in the L. dorsi of the LL group was threefold higher than that of the control group. The leptin mRNA abundance in the L. dorsi of the LL group was 3.3-fold higher than that of the control group ( P < 0.01). These results suggest that a higher activity of adipogenesis may have been involved in the promoted accumulation of IMF in the L. dorsi muscles of pigs, induced by a dietary LL level.
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