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Production and Characterization of Polyclonal Antibody to Transmembrane Inner Ear Protein
YoonYi Nam,Sankarapandian Karuppasamy,Byoungkwon Park,Hyung-Joo Kwon,Jun-Gyo Suh 한국실험동물학회 2009 Laboratory Animal Research Vol.25 No.2
Deafness is one of the most common sensory deficits in human. Since the inner ears of mice and humans are anatomically and physiologically similar, mouse mutants allow us to reveal detailed analysis of defects in the cochlea and vestibular apparatus. Loss of function mutations in transmembrane inner ear (Tmie) protein has been shown to cause deafness in mice (circling, spinner) and humans (DFNB6). However, the detailed mechanism of Tmie protein in the cochlea still remains unclear. First step to achieve that purpose, we produced polyclonal antibody that recognize Tmie protein. The specifically selected amino acid sequence localized at the positions 118-133 (IPGEDKKKKKKDSVD) of Tmie protein has been used as immunogen to produce rabbit polyclonal antibody. Two rabbits were immunized with polypeptide and the serum was harvested from the rabbits served as the source of the polyclonal antibody specific to Tmie. Specificity of the antibody was determined by Western blot analysis. After transfection of tmiePC/pcDNA vector in HEK cells, we identified a specific band of tmie protein. Taken together, our anti-Tmie antibody can be useful for studying localization and characterization of Tmie protein.
Anti-hyperlipidemic effect of soybean extract fermented by Bacillus subtilis MORI in db/db mice
YoonYi Nam,Harry Jung,Sankarapandian Karuppasamy,Jae-Yeon Lee,Kyung-Don Kang,Kyo-Yeol Hwang,Su-Il Seong,Jun-Gyo Suh 한국실험동물학회 2012 Laboratory Animal Research Vol.28 No.2
The purpose of this study was to investigate the anti-hyperlipidemic effect of soy bean extract solution fermented by Bacillus subtilis MORI (BTD-1E) in obese db/db mice. Eight-week-old male db/db mice were administered 33.3 ㎎/㎏ BTD-1E solution orally once a day for four weeks. The BTD-1E group showed significantly lower body weight compared with the db control group (P<0.05). The BTD-1E group showed significantly lower serum total cholesterol and LDL cholesterol levels compared with the db control group, respectively (P<0.05, P<0.01). The BTD-1E group showed significantly decreased liver weight relative to final body weight compared with the db control group (P<0.01). After four weeks of BTD-1E administration, lipid droplets in the liver were apparently decreased in the BTD-1E group compared to the db control group. In summary, our results suggest that BTD-1E has an anti-hyperlipidemic effect in the obese mouse model.
Nam, YoonYi,Kim, Jeong Ki,Cha, Dal-Sun,Cho, Jae-Woo,Cho, Kyu-Hyuk,Yoon, SeokJoo,Yoon, Jong-Bok,Oh, Yang-Seok,Suh, Jun-Gyo,Han, Sang-Seop,Song, Chang-Woo,Yoon, SungJoo Kim Elsevier 2006 Genomics Vol.87 No.4
<P><B>Abstract</B></P><P>A novel autosomal recessive mutant was produced using <I>N</I>-ethyl-<I>N</I>-nitrosourea mutagenesis. The characteristics of the mutant mice included progressive irreversible hair loss within a month of birth, wrinkled skin, and long curved nails. Linkage analysis revealed that the causative gene is linked to <I>D14Mit193</I> on chromosome 14. Sequence analysis of the complete cDNA of the candidate gene, <I>hairless</I> (<I>Hr</I>), identified a homozygous G-to-T transition at nucleotide 3572, leading to the substitution of glycine by tryptophan, designated Gly960Trp. This missense mutation occurs in the vicinity of repression domain 3 of the hairless protein (HR). This allele was named <I>Hr</I><SUP><I>m1Enu</I></SUP><I>.</I> The relative amounts of <I>Hr</I> mRNA and HR protein determined by real-time PCR and Western blot analyses, respectively, were slightly elevated in the mutant mice. Quantitative real-time PCR analysis revealed the increased expression of <I>Kc1</I> and <I>Vdr</I> in the mutant mice, whereas the expression of <I>Nrs1</I> and <I>Krtap16-6</I> was decreased. These results suggest that the Gly960Trp substitution in HR protein in <I>Hr</I><SUP><I>m1Enu</I></SUP> mice may alter the function of HR as a transcriptional corepressor.</P>