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Weihuan Li,Quan Zhang,Xiangqiang Kong,Chunxia Wu,Xiuling Ma,Hui Zhang,Yanxiu Zhao 한국식물학회 2009 Journal of Plant Biology Vol.52 No.2
Vacuolar Na+/H+ antiporters catalyze the exchange of Na+ for H+ across vacuolar membranes and compartmentalize Na+ into vacuoles. They play important roles in cellular pH and Na+ homeostasis. The SsNHX1 gene was previously cloned from a typical euhalophyte, Suaeda salsa. Its cloning revealed another N-terminus truncated transcript, SsNHX2. This potentially alternative splicing variant was truncated from the 49 amino acid residues (aa) of the N terminus of SsNHX1. To compare their degree of salt tolerance, we over-expressed SsNHX1 and SsNHX2 in Arabidopsis. Southern and northern blot analyses showed that both genes had been integrated into that genome and had expressed in several lines. Two types of transgenic plants grew more vigorously than the wild type (WT) under salt stress, but no remarkable differences were found between those SsNHX1 and SsNHX2 transformants. Physiological analyses also supported this phenotype. Both fresh and dry weights of the transgenics as well as their accumulations of Na+ and K+ under salinity were much higher than that of WT, but differences were not significant between SsNHX1 and SsNHX2 plants for any of those parameters. These results suggest that SsNHX2 is a functional Na+/H+ antiporter like SsNHX1 and their levels of salt tolerance are similar.
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( Chao Tong ),( Ning Chen ),( Xun Liao ),( Xuemei Yuan ),( Mengjiao Sun ),( Xiaoliang Li ),( Weihuan Fang ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.9
Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that RecCpp80+10 failed to induce fever in rabbits, whereas RecCpp40<sub>+10</sub> caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.
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