Identification of two novel missense WFS1 mutations,H696Y and R703H,in patients with non-syndromic low-frequency sensorineural hearing loss

Non-syndromic low-frequency sensorineural hearing loss(LFSNHL) is an unusual type of hearing loss in which frequencies≤2000 Hz predominantly are affected.To date,different mutations in two genes,DIAPH1 and WFS1,have been found to be associated with LFSNHL. Here,we report a five-generation Chinese family with postlingual and progressive LFSNHL.We mapped the disease locus to a 2.5 Mb region on chromosome 4p16 between markers SNP_A-2167174 and D4S431,overlapping with the DFNA6/14/38 locus.Sequencing of cand...     

Characterization of barley <Emphasis Type="Italic">Prp1</Emphasis> gene and its expression during seed development and under abiotic stress

The pre-mRNA processing (Prp1) gene encodes a spliceosomal protein. It was firstly identified in fission yeast and plays a regular role during spliceosome activation and cell cycle. Plant Prp1 genes have only been identified from rice, Sorghum and Arabidopsis thaliana. In this study, we reported the identification and isolation of a novel Prp1 gene from barley, and further explored its expressional pattern by using real-time quantitative RT-PCR, promoter prediction and analysis of microarray data. The putative barley Prp1 protein has a similar primary structure features to those of other known Prp1 protein in this family. The results of amino acid comparison indicated that Prp1 protein of barley and other plant species has a highly conserved 3′ termnal region while their 5′ sequences greatly varied. The results of expressional analysis revealed that the expression level of barley Prp1 gene is always stable in different vegetative tissues, except it is up-regulated at the mid- and late stages of seed development or under the condition of cold stress. This kind of expressional pattern for barley Prp1 is also supported by our results of comparison of microarray data from barley, rice and Arabidopsis. For the molecular mechanism of its expressional pattern, we conclude that the expression of Prp1 gene may be up-regulated by the increase of pre-mRNAs and not be constitutive or ubiquitous.     

Molecular cloning and characterization of caprine <Emphasis Type="Italic">calpastatin</Emphasis> gene

Calpastatin (CAST) is an important gene for meat quality traits in livestock and poultry. The cDNA of caprine CAST gene was amplified for the first time using RACE-PCR. Results showed the full-length cDNA of caprine CAST gene (Accession no. GU944861) was 2435 base pair (bp) and contained a 2187 bp open reading frame encoding a protein with 728 amino acid residues. Bioinformatic analysis indicated that caprine CAST cDNA was 89.8–95.4, 83.5–92.2, 72.8–81.8 and 69.8–73.5% identical to sheep, cattle, pig and human CAST cDNA. It was predicted that caprine CAST contained four conserved domains with 42 serine phosphorylation loci, 18 threonine phosphorylation loci, 1 tyrosine phosphorylation locus and 5 specific PKC phosphorylation loci. This work provided an important experimental basis for further research on the function of CAST in goat.     

Derivation and characterization of ovine embryonic stem-like cell lines in semi-defined medium without feeder cells

Domestic animal embryonic stem (ES) cells would provide an invaluable research tool for genetic breeding and the production of transgenic animals. Unfortunately, authentic domestic animals ES cells have not been established despite progress made over more than two decades. Here, we show that ovine ES-like cells can be efficiently derived and propagated in a semi-defined medium that contains N2, B27, GSK3 inhibitor (CHIR99021), and basic fibroblast growth factor (bFGF). These ovine ES-like cells had a characteristic three-dimensional appearance, showed a bFGF dose-dependence, expressed specific markers such as alkaline phosphatase (AP), Oct-4, Sox2, Nanog and can be maintained for 30 passages. Moreover, these cells differentiated in vitro into neuronal cells, and formed teratomas containing a variety of different tissues including cartilage and neural tissue when injected into kidney capsules of severe combined immunodeficiency (SCID) mice. But the cell lines fail to contribute to embryonic development upon blastocyst transplantation. To our knowledge, this is the first experiment to use semi-defined medium without feeder-cells to derive ES-like cells from ovine blastocysts, and opens the door to deriving authentic ES cells from domesticated ungulates.     

Piezoelectric urea biosensor based on immobilization of urease onto nanoporous alumina membranes

The urease was immobilized onto nanoporous alumina membranes prepared by the two-step anodization method, and a novel piezoelectric urea sensing system with separated porous alumina/urease electrode has been developed through measuring the conductivity change of immobilized urease/urea reaction. The process of urease immobilization was optimized and the performance of the developed urea biosensor was evaluated. The obtained urea biosensor presented high-selectivity monitoring of urea, better reproducibility (S.D. = 0.02, n = 6), shorter response time (30 s), wider linear range (0.5 μM to 3 mM), lower detection limit (0.2 μM) and good long-term storage stability (with about 76% of the enzymatic activity retained after 30 days). The clinical analysis of the urea biosensor confirmed the feasibility of urea detection in urine samples.     

Taxonomy and phylogeny of Ceriporia (Polyporales,Basidiomycota) with an emphasis of Chinese collections

Ceriporia accommodates a kind of wood-inhabiting polypores producing resupinate basidiocarps and causing a white rot. More than 30 species of this genus have been described; however, only a few species were referred to molecular phylogeny. In this study, a total of 203 specimens of Ceriporia were studied morphologically, and the ITS and/or nLSU regions from 42 samples, representing 18 species, were sequenced for phylogenetic analysis. Based on both morphological and phylogenetic analyses, three new species of Ceriporia, C. bubalinomarginata, C. pseudocystidiata and C. variegata, are described and illustrated. An annotated identification key is provided for all 20 species of this genus thus far known in China. Our phylogeny shows that (1) Ceriporia is not monophyletic, (2) C. spissa and C. viridans as morphologically circumscribed are polyphyletic, (3) C. inflata is retained for both C. inflata and C. jiangxiensis, and (4) presence or absence of hymenial cystidia is not a useful character in delimiting species relationships in Ceriporia.