摘要：以含有绵羊叠朊编码基因Prnd的ORF的质粒OvPrnd-T为模板,PCR扩增出成熟叠朊编码基因片段,并将其插入原核表...以含有绵羊叠朊编码基因Prnd的ORF的质粒OvPrnd-T为模板,PCR扩增出成熟叠朊编码基因片段,并将其插入原核表达载体pET-30a(+)的EcoRⅠ和XhoⅠ位点之间,构建出重组表达质粒pET-OvmDpl.将pET-OvmDpl电转化到宿主菌株BL21(DE3)中,以1.0 mM IPTG诱导表达,SDS-PAGE结果显示绵羊成熟叠朊被大肠杆菌表达.以Ni-NTA树脂纯化表达产物,电泳结果显示纯化产物中存在与预计表达蛋白分子量23Ku二倍相当的蛋白条带,分析该蛋白可能是成熟叠朊在大肠杆菌表达时形成的二聚体结构.显示全部
摘要： TOXOPLASMA GONDII,AN OBLIGATE INTRACELLULAR PARASITE,CAN INFECT ALMOST ALL MAMMALIAN SPECIES AN... TOXOPLASMA GONDII,AN OBLIGATE INTRACELLULAR PARASITE,CAN INFECT ALMOST ALL MAMMALIAN SPECIES AND MAY LEAD TO TOXOPLASMOSIS IN THESE HOSTS.IN THE PRESENT STUDY,WE EXAMINED SEQUENCE VARIATION IN RHOPTRY PROTEIN 38 (ROP38) GENES AMONG T.GONDII ISOLATES FROM DIFFERENT HOSTS AND GEOGRAPHICAL REGIONS.THE COMPLETE ROP38 GENE FROM 13 T.GONDII ISOLATES WERE AMPLIFIED AND SEQUENCED.THE RESULTS OF SEQUENCE ALIGNMENTS SHOWED THAT THE LENGTHS OF THE ENTIRE ROP38 GENE RANGED FROM 2646 BP TO 2650 BP,AND THE SEQUENCE VARIATION WAS 0:1.1% AMONG THE 13 T.GONDII ISOLATES,INDICATING A LOW SEQUENCE VARIATION IN ROP38 GENE.PHYLOGENETIC ANALYSIS OF ROP38 SEQUENCES USING BAYESIAN INFERENCE (BI) SHOWED THAT THE CLUSTERING OF THE 13 T.GONDII ISOLATES WERE NOT CONSISTENT WITH THEIR RESPECTIVE GENOTYPES,INDICATING THAT THE ROP38 GENE IS NOT A SUITABLE GENETIC MARKER FOR POPULATION GENETIC STUDIES OF DIFFERENT T.GONDII GENOTYPES FROM DIFFERENT HOSTS AND GEOGRAPHICAL LOCATIONS,BUT MAY REPRESENT AN POTENTIAL VACCINE CANDIDATE AGAINST TOXOPLASMOSIS,WORTH FURTHER STUDIES.显示全部
摘要： ANALYSES OFMITOCHONDRIAL (MT) GENOME SEQUENCES IN RECENT YEARS CHALLENGE THE CURRENT WORKING HY... ANALYSES OFMITOCHONDRIAL (MT) GENOME SEQUENCES IN RECENT YEARS CHALLENGE THE CURRENT WORKING HYPOTHESIS OF NEMATODA PHYLOGENY PROPOSED FROM MORPHOLOGY,ECOLOGY AND NUCLEAR SMALL SUBUNIT RRNA GENE SEQUENCES,AND RAISE THE NEED TO SEQUENCE ADDITIONAL MT GENOMES FOR A BROAD RANGE OF NEMATODE LINEAGES.WE SEQUENCED THE COMPLETE MT GENOMES OF THREE ASCARIDIA SPECIES,NAMELY A.GALLI,A.COLUMBAE AND ASCARIDIA SP.(GHL:2012),THAT INFEST CHICKENS,PIGEONS AND PARROTS,RESPECTIVELY.THESE THREE ASCARIDIA SPECIES HAVE AN IDENTICAL ARRANGEMENT OFMT GENES TO EACH OTHER BUT DIFFER SUBSTANTIALLY FROM OTHER NEMATODES.PHYLOGENETIC ANALYSES OF THE MT GENOME SEQUENCES OF THE ASCARIDIA SPECIES,TOGETHER WITH 62 OTHER NEMATODE SPECIES,SUPPORT THE MONOPHYLIES OF SEVEN HIGH:LEVEL TAXA OF THE PHYLUM NEMATODA: 1) THE SUBCLASS DORYLAIMIA; 2) THE ORDERS RHABDITIDA,TRICHINELLIDA AND MERMITHIDA; 3) THE SUBORDER RHABDITINA; AND 4) THE INFRAORDERS SPIRUROMORPHA AND OXYURIDOMORPHA.ANALYSES OFMT GENOME SEQUENCES,HOWEVER,REJECT THE MONOPHYLIES OF THE SUBORDERS SPIRURINA AND TYLENCHINA,AND THE INFRAORDERS RHABDITOMORPHA,PANAGROLAIMOMORPHA AND TYLENCHOMORPHA.MONOPHYLY OF THE INFRAORDER ASCARIDOMORPHA VARIES DEPENDING ON THE METHODS OFPHYLOGENETIC ANALYSIS.THE ASCARIDOMORPHA WAS MORE CLOSELY RELATED TO THE INFRAORDERS RHABDITOMORPHA AND DIPLOGASTEROMORPHA (SUBORDER RHABDITINA) THAN THEY WERE TO THE OTHER TWO INFRAORDERS OF THE SPIRURINA: OXYURIDORPHA AND SPIRUROMORPHA.THE CLOSER RELATIONSHIP AMONG ASCARIDOMORPHA,RHABDITOMORPHA AND DIPLOGASTEROMORPHA WAS ALSO SUPPORTED BY A SHARED COMMON PATTERN OFMITOCHONDRIAL GENE ARRANGEMENT.ANALYSES OF MITOCHONDRIAL GENOME SEQUENCES AND GENE ARRANGEMENT HAS PROVIDED NOVEL INSIGHTS INTO THE PHYLOGENETIC RELATIONSHIPS AMONG SEVERAL MAJ OR LINEAGES OF NEMATODES.MANY LINEAGES OF NEMATODES,HOWEVER,ARE UNDERREPRESENTED OR NOT REPRESENTED IN THESE ANALYSES.EXPANDING TAXON SAMPLING IS NECESSARY FOR FUTURE PHYLOGENETIC STUDIES OF NEMATODES WITH MT GENOME SEQUENCES.显示全部
摘要： THIS STUDY EXAMINED SEQUENCE DIFFERENCES IN MITOCHONDRIAL CYTOCHROME C OXIDASE SUBUNIT(COX1),LA... THIS STUDY EXAMINED SEQUENCE DIFFERENCES IN MITOCHONDRIAL CYTOCHROME C OXIDASE SUBUNIT(COX1),LARGE SUBUNIT RIBOSOMAL RNA (RRNL) AND NADH DEHYDROGENASE SUBUNITS 1 AND 4 (NADL AND NAD4) BETWEEN CHABERTIA OVINA AND C.ERSCHOWI FROM YAKS IN QINGHAI AND GOATS IN SHAANXI PROVINCES,CHINA.A PART OF THE COX 1 (PCOX 1),RRNL (PRRNL),NAD1 AND NAD4 GENES (PNAD1 AND PNAD4) WERE AMPLIFIED SEPARATELY FROM INDIVIDUAL NEMATODES BY PCR AND SEQUENCED.THE LENGTH OF THE SEQUENCES OFPCOX1,PRRNL,PNADL AND PNAD4 WAS 441 BP,450 BP,526 BP AND 914 BP FOR C.OVINA,AND 441 BP,451 BP,?517 BP AND 810 BP FOR C.ERSCHOWI,RESPECTIVELY.THE INTRA:SPECIFIC SEQUENCE VARIATIONS WITHIN C.OVINA WERE 0.2%:2.9% FOR PCOX1,0:0.9% FOR PRRNL,0.6%:2.3% FOR PNAD1,AND 0.4%:2.0% FOR PNAD4,AND WERE 0.5%: 1.6% FOR PCOX1,0:1.1% FOR PRRNL,0.2%: 1.7% FOR PNAD1,AND 0.4%: 1.1% FOR PNAD4 WITHIN C.ERSCHOWI.WHEREAS,THE INTER:SPECIFIC SEQUENCE DIFFERENCES BETWEEN THE TWO SPECIES WERE OBVIOUSLY HIGHER,BEING 11.6%: 12.9% FOR PCOX1,9.8%: 11.1% FOR PRRNL,14.4%: 15.9% FOR PNADL,AND 16.4%: 17.7% FOR PNAD4.PHYLOGENETIC ANALYSES USING BAYESIAN INFERENCE (BI),BASED ON COMBINED SEQUENCES OF FOUR GENES,INDICATED THAT THE C.OVINA AND C.ERSCHOWI REPRESENT DISTINCT SPECIES.THESE RESULTS DEMONSTRATE THAT THESE MT GENE SEQUENCES PROVIDE NOVEL GENETIC MARKERS FOR THE IDENTIFICAITON AND DIFFERENTIATION C.OVINA AND C.ERSCHOWI,AND HAVE IMPLICATIONS FOR STUDYING THE POPULATION GENETICS AND MOLECULAR EPIDEMIOLOGY OF CHABERTIA SPP.显示全部
