Cloning and characterization of nanos gene in silkworm Bombyx mori

Gene nanos is a maternal posterior group gene required for normal development of abdominal segments and the germ line in Drosophila. Expression of nanos-related genes is associated with the germ line in a broad variety of other taxa. In this study, the 5＇-RACE method and the in silico cloning method are used to isolate the new nanos-like gene of Bombyx mor/and the gene obtained is analyzed with bioinformatics tools. The putative protein is expressed in Escherichia coli and the antiserum has been produced in New Zealand white rabbits. The result shows that the nanos cDNA is 1,913 bp in full length and contains a 954 bp open reading frame. The deduced protein has 317 amino acid residues, with a predicted molecular weight of 35 kDa, isoelectric point of 5.38, and contains a conserved nanos RNA binding domain. The conserved region of the deduced protein shares 73% homology with the nanos protein conserved region of Honeybee （Apis mellifera）. This gene has been registered in the GenBank under the accession number EF647589. One encoding sequence of the nanos fragment has been successfully expressed in E. coli. Western blotting analysis indicates that homemade antiserum can specifically detect nanos protein expressed in prokaryotic cells.

Comparison of electrocardiogram characteristics between left bundle branch pacing and right ventricular septal pacing in patients receiving pacemaker therapy

Background and Objective Cardiac pacing is an effective therapy in patients with bradycardia.Conventional right ventricular(RV)pacing is the source of ventricular dyssynchrony,leading to unfavorable clinical outcome.This study compared the electrocardiogram(ECG)characteristics during left bundle branch pacing(LBBP)with that during RV septal pacing(RVSP)which has been thought to be better than RV apical pacing.

A robust protein extraction method for two dimensional electrophoresis of silkworm proteins

Silkworm (Bombyx mori) is a species of agricultural importance, as well as a model organism for Lepidoptera insects. Proteomic method has been widely used in silkworm research, and a robust mass spectrometry-compatible protein extraction method is urgently needed. In this study, we adapted phenol extraction method to extract silkworm midgut protein, and coupled this method with pH 5 - 8 gel strip for two dimensional electrophoresis. The phenol extraction method significantly increased the resolution, as well as greatly reduced the background of two dimensional electrophoresis gels. In addition, this method was well compatible with mass spectrometry analysis. This is the first report that phenol extraction method is used for silkworm midgut protein extraction, and may be applied in other researches.

Molecular cloning, expression and characterization of a novel geneβ-N-acetylglucosaminidase from Bombyxmori

Previously, we have reported that a gene encoding Bombyxmoriβ-N-acetylglucosaminidase 2 (BmGlcNAcase2) has been identified differentially expressed in the midgut of Bombyxmori strain NB resistant to nucleopolyhedrovirus (BmNPV), strain 306 susceptible to NPV and a near isogenic line BC9 with similar genetic background to 306 but resistant to NPV by two-dimensional gel electrophoresis (2-DE). To get more knowledge about the relationship between β-N-acetylglucosaminidase and the resistance of NPV, in this study, the 1542 bp open reading frame of a putative bombyxmoriβ-N-acetylglucosaminidase 2 gene (BmGlcNAcase2) was amplified from a pool of bombyxmoricDNAs and inserted into the prokaryotic expression plasmid pET-30a(+).Western blotting analysis showed that BmGlcNAcase2 was expressed in hemolymph, ovary, testis, fat body, trachea, midgut and silk gland of fifth instar larvae respectively . Immunofluoresence analysis indicated that BmGlcNAcase2 was mainly located to the cytoplasm or some structure in cytoplasm.

Analysis of the Hydraulic and Heat Transfer Evolution Mechanism of a Single Rock Fracture

Mineral dissolution and mechanical deformation of granite are two main mechanisms that affect permeability evolution of rock fracture.In this study,two water flow-through experiments with large granite fractures were conducted at 200 0C with a constant flow rate for 24 h,under confining pressures of 5 and 10 MPa,respectively.Water pressure and temperature were measured,fracture aperture and permeability were calculated,and chemical element concentrations in effluent water were tested for mechanism analysis.The permeability fluctuates up and down between 2.62 × 10^(-12)and 3.16 ×10^(-12)m^(2)at confining pressure of 5 MPa;while it decreased monotonously by 24% from 1.92 × 10^(-12)to1.45 × 10^(-12)m^(2)at a confining pressure of 10 MPa.The heat transfer rates at both experiments stay stable at about 0.25 J/s.The mass concentration of Ca,Na,K,and Si in effluent water are between 5 to 23mg/L,indicating slight dissolution of Ca-plagioclase,Na-plagioclase,and K-feldspar,as well as possible precipitation of minor amount of kaolinite or quartz.The total amount of free-face dissolution and pressure dissolution are similar at 5 and 10 MPa.The geochemical reaction counts for only small part of the aperture change,while the mechanical deformation counts the major part of the aperture change.

Comparison of immediate changes of repolarization parameters after left bundle branch area pacing and traditional biventricular pacing in heart failure patients

To the Editor:Cardiac resynchronization therapy(CRT)is an effective treatment for advanced chronic heart failure(HF)patients with reduced ejection fraction.Despite its beneficial effects on cardiac function,traditional biventricular epicardial pacing(BiVP-epi)could not reduce ventricular arrhythmias(VAs)burden in all HF patients.

Functional analysis of zona pellucida domain protein Dusky in Tribolium castaneum

The zona pellucida domain protein Dusky(Dy)plays a vital role in wing morphogenesis in insects,but little information on its function has been reported.In this study,we found that dy regulated wing cell size,larval and pupal duration,and the metabolism of amino acid and 20-hydroxyecdysone in Tribolium castaneum.Using RNA-seq,413 differentially expressed genes were identified between physiological buffer-injected and dy-double-stranded RNA-treated larvae,including 88 downregulated genes and 325 upregulated genes.Among these genes,dy knockdown increased CYP18A1 expression to elevate the 26-hydroxylation of 20-hydroxyecdysone,which ultimately led to growth defects in wing cells.Silencing of dy upregulated the transcription of genes encoding tyrosine aminotransferase,4-hydroxyphenylpyruvate dioxygenase,homogentisate 1,2-dioxygenase,and Pale to promote the catabolism of tyrosine and phenylalanine,which eventually reduced amino acid content.Furthermore,dy knockdown upregulated 4E-BP expression,and 4E-BP silencing partially phenocopied dy RNA interference-mediated wing morphogenesis.These results suggest that Dy controls 20-hydroxyecdysone and amino acid metabolism to regulate wing morphogenesis in the insect.