西藏牦牛ADD1基因第2外显子的PCR-SSCP检测及序列分析

利用PCR-SSCP和DNA测序方法对帕里牦牛、工布江达牦牛、类乌齐牦牛、康布牦牛、桑日牦牛、巴青牦牛、江达牦牛、斯布牦牛、嘉黎牦牛、桑桑牦牛、丁青牦牛等西藏11个牦牛类群共483头牦牛的ADD1基因(被认为是极可能影响肉质的候选基因)第2外显子序列进行遗传多态性分析。结果表明,66 bp处碱基T缺失和256 bp处A→G突变,共有AA、AB、BB 3种基因型;其中帕里牦牛只有AA基因型,江达牦牛只有AB基因型,康布牦牛、嘉黎牦牛、巴青牦牛、桑日牦牛、斯布牦牛均缺少BB基因型,丁青牦牛、类乌齐牦牛缺少AB型,均存在严重偏态;桑桑牦牛、工布江达牦牛中存在AA、AB、BB 3种基因型。除江达牦牛外,其它10个牦牛类群中AA为优势基因型,A基因为优势等位基因,分布较高。除帕里牦牛、巴青牦牛外,其它9个牦牛类群均处于中度多态(0.25相似文献   

An α-glucan isolated from root of Isatis Indigotica,its structure and adjuvant activity

The root of Isatis indigotica is a traditional Chinese herbal medicine. An α-glucan (IIP-A-1) was firstly isolated from the roots. In this study we elucidated the chemical structure of IIP-A-1 and determined its adjuvant activity by co-immunizing mice with H1N1 influenza virus split and recombinant hepatitis B surface antigen (HBsAg), respectively. The polysaccharide was pretreated with periodate oxidation, Smith degradation and methylation in order to analyze its structure using GC, HPGPC, FT-IR, NMR and GC-MS. The adjuvant effect was evaluated by determining the antibody titers of serum against H1N1 influenza and HBsAg using ELISA. The proliferation and TNF-α secretion of macrophages administrated with different dose of IIP-A-1 were measured in vitro. The results of this study revealed that IIP-A-1 was an α-glucan with the molecular weight of 3,600 Da. The backbone was α-(1?→?4)-D-glucan with (1?→?6) branch chain. The α-glucan could significantly enhance the immune response of mice immunized with H1N1 influenza or HBsAg in vivo and exert good dose-dependent effects on the proliferation and the TNF-α secretion of macrophages in vitro. These results supported that IIP-A-1 was expected to be an efficacious adjuvant candidate for prophylactic and therapeutic vaccines.     

Effect of biochar amendment on maize yield and greenhouse gas emissions from a soil organic carbon poor calcareous loamy soil from Central China Plain

Aims

A field experiment was conducted to investigate the effect of biochar on maize yield and greenhouse gases (GHGs) in a calcareous loamy soil poor in organic carbon from Henan, central great plain, China.

Methods

Biochar was applied at rates of 0, 20 and 40?t?ha^?1 with or without N fertilization. With N fertilization, urea was applied at 300?kg?N ha^?1, of which 60% was applied as basal fertilizer and 40% as supplementary fertilizer during crop growth. Soil emissions of CO₂, CH₄ and N₂O were monitored using closed chambers at 7?days intervals throughout the whole maize growing season (WMGS).

Results

Biochar amendments significantly increased maize production but decreased GHGs. Maize yield was increased by 15.8% and 7.3% without N fertilization, and by 8.8% and 12.1% with N fertilization under biochar amendment at 20?t?ha^?1 and 40?t?ha^?1, respectively. Total N₂O emission was decreased by 10.7% and by 41.8% under biochar amendment at 20?t?ha^?1 and 40?t?ha^?1 compared to no biochar amendment with N fertilization. The high rate of biochar (40?t?ha^?1) increased the total CO₂ emission by 12% without N fertilization. Overall, biochar amendments of 20?t?ha^?1 and 40?t?ha^?1 decreased the total global warming potential (GWP) of CH₄ and N₂O by 9.8% and by 41.5% without N fertilization, and by 23.8% and 47.6% with N fertilization, respectively. Biochar amendments also decreased soil bulk density and increased soil total N contents but had no effect on soil mineral N.

Conclusions

These results suggest that application of biochar to calcareous and infertile dry croplands poor in soil organic carbon will enhance crop productivity and reduce GHGs emissions.     

Guanylate kinase domains of the MAGUK family scaffold proteins as specific phospho-protein-binding modules

Membrane-associated guanylate kinases (MAGUKs) are a large family of scaffold proteins that play essential roles in tissue developments, cell-cell communications, cell polarity control, and cellular signal transductions. Despite extensive studies over the past two decades, the functions of the signature guanylate kinase domain (GK) of MAGUKs are poorly understood. Here we show that the GK domain of DLG1/SAP97 binds to asymmetric cell division regulatory protein LGN in a phosphorylation-dependent manner. The structure of the DLG1 SH3-GK tandem in complex with a phospho-LGN peptide reveals that the GMP-binding site of GK has evolved into a specific pSer/pThr-binding pocket. Residues both N- and C-terminal to the pSer are also critical for the specific binding of the phospho-LGN peptide to GK. We further demonstrate that the previously reported GK domain-mediated interactions of DLGs with other targets, such as GKAP/DLGAP1/SAPAP1 and SPAR, are also phosphorylation dependent. Finally, we provide evidence that other MAGUK GKs also function as phospho-peptide-binding modules. The discovery of the phosphorylation-dependent MAGUK GK/target interactions indicates that MAGUK scaffold-mediated signalling complex organizations are dynamically regulated.     

Expression, purification, and activity assay of peptide deformylase from Escherichia coli and Staphylococcus aureus

Peptide deformylase (PDF) is considered an attractive target for screening novel antibiotics. The PDF from Escherichia coli and Staphylococcus aureus are representative of the gram-negative species type of PDF (type I PDF) and the gram-positive species type of PDF (type II PDF), respectively. They could be used for screening broad-spectrum antibiotics. Herein, we cloned the def gene by PCR, inserted it into plasmid pET-22b-def, and transformed the plasmid into E. coli BL21 (DE3) cells, then the cells were induced by IPTG to express PDF. E. coli Ni(2+)-PDF was extracted and purified by ion-exchange chromatography and gel filtration chromatography. S. aureus PDFs were extracted and purified using the MagExtractor kit. The nickel form of S. aureus PDF was obtained by adding NiCl(2) to all reagents used for purification. Iron-enriched S. aureus PDF was obtained by adding FeCl(3) to the growth medium for E. coli BL21 (DE3) cells and adding FeCl(3) and catalase to all reagents used for purification. The activities of PDFs were analyzed, compared, and grouped according to the experimental conditions that produced optimal activity, and we used actinonin as an inhibitor of PDF and calculated the IC(50) value. We obtained high expression of E. coli and S. aureus PDF with high activity and stability. The function of PDFs was inhibited by actinonin in a dose-dependent manner. Results may be helpful for future mechanistic investigations of PDF as well as high-throughput screening for other PDF inhibitors.     

Mutational analysis of the role of GXXXG motif in the function of human organic anion transporter 1 (hOAT1)

Human organic anion transporter hOAT1 plays a critical role in the body disposition of environmental toxins and clinically important drugs including anti-HIV therapeutics, anti-tumor drugs, antibiotics, anti-hypertensives, and anti-inflammatories. hOAT1 has two GXXXG motifs in its transmembrane domains 2 and 5, a motif linked to the protein processing and oligomerization of other proteins. In the current study, we substituted glycine of these GXXXG motifs with alanine and evaluated the effect of such mutations on the expression and function of hOAT1. Mutations of GXXXG motif in the transmembrane domain 2 resulted in mutants G144A and G148A, both of which had no transport activity due to complete loss in the surface and total cell expression of the transporter protein. Treatment of G144A- and G148A-expressing cells with proteasomal inhibitor resulted in the recovery of ER-resident immature form of hOAT1, but not its surface-resident mature form, whereas treatment of these cells with lysosomal inhibitor had no effect on the expression of the mutant transporters. Mutations of GXXXG motif in the transmembrane domain 5 resulted in mutants G223A and G227A, among which only G227 had dramatic reduction of transport activity due to dramatic loss in the surface and total cell expression of the transporter. The reduction in the surface expression of G227 was consistent with the decrease in maximum transport velocity Vmax. Treatment of G227A-expressing cells with proteasomal inhibitor or lysosomal inhibitor resulted in partial recovery of both the immature form and the mature form of hOAT1 in the total cell extracts. However, such partial recovery of the mature form in total cell extracts did not lead to the partial recovery of surface expression and function of the transporter. Our data suggest that the GXXXG motifs in transmembrane domains 2 and 5 play critical roles in the stability of hOAT1.     

Cold effect analysis and screening of SNPs associated with cold‐tolerance in the olive flounder Paralichthys olivaceus

Olive flounder Paralichthys olivaceus is an important mariculture fish and cold stress limits to its growth and survival during winter. In the present study, the cold‐tolerant (CT) and cold‐sensitive (CS) flounder were defined after cold treatment (0.7 ± 0.05°C). Cold effects on histological and physiological and gene expression levels were analyzed. The correlation analyses between single nucleotide polymorphism locus (SNP) in hsp70, yb‐1 and hmgb1 and the trait of cold tolerance were performed. The results showed that flounder gill was obviously damaged after treatment and more serious lesions in some of the epithelial cell layers were observed in the CS group. ATP concentration and pyruvate kinase activity in the CT group were significantly higher than those in the CS group (p < 0.05). Expression levels of hsp70 and yb‐1 were both up‐regulated markedly after cold treatment (p < 0.05). Nine, twenty‐one and ten SNPs were screened from hsp70, yb‐1 and hmgb1 partial sequences, respectively. Among them, 3 SNPs in hsp70 and 2 SNPs in hmgb1 showed significant difference in the two groups. Allele G of the SNP 8 (locus 1797) in hsp70 was only observed in the CT group. In the SNP 7 (locus 725) of hmgb1, genotype TT and allele T, and genotype CC and allele C were associated with cold tolerance and cold sensitive, respectively. Furthermore, one haplotype (TTG) generated from SNPs of hsp70 gene among CT and CS individuals showed significant relationship with cold tolerance (p = 0.031). Two haplotypes (ATG) and (TCT) generated from SNPs of hmgb1 were significantly associated with resistance (p = 0.001) and sensitive (p = 0.002) in cold treatment, respectively. These SNPs, genotype, alleles and haplotypes of hsp70 and hmgb1 may be related to the cold resistance of flounder, and could be candidate markers potentially for further selective breeding.     

Two new species and five newly recorded species of the genus Udea Guenée from China (Lepidoptera,Crambidae)

A checklist of the 31 Chinese species of Udea is given, including the new species and new records. Udea curvata sp. n. and Udea albostriata sp. n. are described and illustrated. Udea exigualis (Wileman, 1911), Udea stationalis Yamanaka, 1988, Udea prunalis (Denis & Schiffermüller, 1775), Udea elutalis (Denis & Schiffermüller, 1775) and Udea cyanalis (La Harpe, 1855) are newly recorded for China.