In this study, we investigated suitable conditions for the reduction of disulfides in mouse serum samples by tris(2-carboxyethyl) phosphine (TCEP) for fast analysis of total biothiols. Disulfides were reduced with TCEP, and then, thiols were derivatized with the fluorogenic reagent, ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate (SBD-F). Interference peaks on chromatograms of mouse serum samples disappeared when the TCEP reaction was conducted on ice instead of at room temperature, which is used classically. Low-molecular-weight disulfides, such as cystine and glutathione disulfide, were nearly completely reduced by TCEP on ice. Six SBD-biothiols (homocysteine, cysteine, cysteinylglycine, glutathione, γ-glutamylcysteine, and
N
-acetylcysteine) were separated within 7.5 min on a sulfoalkylbetain-type column (ZIC-HILIC: 150 × 2.1 mm i.d., 3.5 μm), without interference peaks. The developed method showed good linearity and reproducibility, with inter- and intra-day precisions of less than 3%.
Rong-qing, G., Lan-ping, W., Hong, C., Yong-hong, Li., De-jun, J. and Chun-fang, C. 2009. Variations in the promoter region, the first exon and part of the first intron of bovine growth hormone gene. J. Appl. h i m . Res., 35: 155-158.Variations i n the promoter region, the first exon and part of the first intron of bovine growth hormone (bGH) gene were identified from five indigenous Chinese bovine species. Cloned and sequenced amplified fragments revealed two different bands (450bp and 453bp) compared with the GenBank bGH sequence M57764. Two new SNPs in Leiqiong cattle and one SNP in Gayal were identified in the promoter region. Four new SNPs were identified i n Leiqiong cattle and one both from Mongolian cattle and Bazhou yak at the same position within the first intron. Regulatory elements of TATA box, CRE, NRE3, dPitl and pPitl were also identified in the promoter region of GH in five bovine species, suggesting to be strictly conservative during the course of evolution.
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