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31.
32.
韩玉珉 《中国生物化学与分子生物学报》1990,6(1):66-70
用寡聚核苷酸诱导的定位突变法,将人U_1和U_2snRNA基因的5'-端调控区域的一段能与SV_(40)T抗原相结合的DNA删去,造成缺失突变,改变这段DNA核苷酸的排列顺序,造成取代突变。突变株用原位杂交法筛选,由限制性内切酶电泳图谱分析和DNA顺序测定得到证实。突变率约为5%。 相似文献
33.
34.
Associations between different agonistic and affiliative behavioural patterns of female domestic cats (Felis silvestris catus) were studied. In three groups of intact cats living in confinement frequencies of fourteen agonistic and affiliative behavioural
patterns were recorded. The technique of factor analysis (Principal Components Analysis followed by varimax rotation on a
dyads X behavioural patterns matrix) was used to detect clusters in these behavioural patterns. Five factors (or types of
interindividual relationships) were extracted per group. They accounted collectively for at least 77% of the total variance
present in the data. Although differences existed between groups with respect to behavioural patterns included in each factor,
four clusters of behaviours could be discriminated: (I) social rubbing, lordosis and rolling in front of partner (sexual behaviour),
(II) allogrooming, social sniffing, nosing, sniffing rear and treading (inspection-affiliative behaviour), (III) offensive
behaviour and staring, and (IV) defensive behaviour and staring. The role of these clusters in group living is discussed. 相似文献
35.
Use of the additive main effects and multiplicative interaction model in QTL mapping for adaptation in barley 总被引:4,自引:0,他引:4
Ignacio Romagosa Steven E. Ullrich Feng Han Patrick M. Hayes 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,93(1-2):30-37
The additive main effects and multiplicative interaction (AMMI) model has emerged as a powerful analytical tool for genotype x environment studies. The objective of the present study was to assess its value in quantitative trait locus (QTL) mapping. This was done through the analysis of a large two-way table of genotype-by-environment data of barley (Hordeum vulgare L.) grain yields, where the genotypes constituted a genetic population suitable for mapping studies. Grain yield data of 150 doubled haploid lines derived from the Steptoe x Morex cross, and the two parental lines, were taken by the North American Barley Genome Mapping Project (NABGMP) at 16 environments throughout the barley production areas of the USA and Canada. Four regions of the genome were responsible for most of the differential genotypic expression across environments. They accounted for approximately 50% of the genotypic main effect and 30% of the genotype x environment interaction (GE) sums of squares. The magnitude and sign of AMMI scores for genotypes and sites facilitate inferences about specific interactions. The parallel use of classification (cluster analysis of environments) and ordination (principal component analysis of GE matrix) techniques allowed most of the variation present in the genotype x environment matrix to be summarized in just a few dimensions, specifically four QTLs showing differential adaptation to four clusters of environments. Thus, AMMI genotypic scores, when the genotypes constituted a population suitable for QTL mapping, could provide an adequate way of resolving the magnitude and nature of QTL x environment interactions.Ignacio Romagosa was on sabbatical leave from the University of Lleida and the Institut de Recerca i Tecnologia Agroalimentàries, Lleida, Spain, when this study was conducted 相似文献
36.
K Han 《Nucleic acids research》1996,24(21):4362-4363
I have developed an efficient method for transfecting Drosophila S2 cells using DDAB, a cationic liposome reagent. The optimized DDAB method resulted in a 10 times or greater increase in transfection efficiency compared with the conventional calcium phosphate method which has been essentially the only way for transfecting S2 cells. 相似文献
37.
重组水蛭素HV2的稳定性 总被引:3,自引:0,他引:3
重组水蛭素HV2是凝血酶的特异性抑制剂,是一种非常稳定的蛋白质。温度的升高(100℃水浴)和pH(1─13)的改变不影响其活力,在某些变性剂(8mol/L尿素、1%SDS和6mol/L盐酸胍)存在的条件下也非常稳定,0.1mol/L的DTT在70℃时使其部分失活,只有pH和温度同时升高其活力才开始下降,pH13、80℃处理15min即完全失活,氨基酸组成和活性分析发现失活样品的Cys和Lys被破坏。重组水蛭素HV2含有一个结构紧密的N端核心区和一个无序的C端尾部。其N端的3个Lys-Xaa键均不被胰蛋白酶水解;胃蛋白酶及糜蛋白酶消化后,分离所得片段,氨基酸组成分析发现N端核心区依然保持很高的抗凝血酶活性,继续消化24h,核心区不被进一步降解。 相似文献
38.
Identification of residues of the H-ras protein critical for functional interaction with guanine nucleotide exchange factors. 总被引:7,自引:7,他引:0 下载免费PDF全文
Ras proteins are activated in vivo by guanine nucleotide exchange factors encoded by genes homologous to the CDC25 gene of Saccharomyces cerevisiae. We have taken a combined genetic and biochemical approach to probe the sites on Ras proteins important for interaction with such exchange factors and to further probe the mechanism of CDC25-catalyzed GDP-GTP exchange. Random mutagenesis coupled with genetic selection in S. cerevisiae was used to generate second-site mutations within human H-ras-ala15 which could suppress the ability of the Ala-15 substitution to block CDC25 function. We transferred these second-site suppressor mutations to normal H-ras and oncogenic H-rasVal-12 to test whether they induced a general loss of function or whether they selectively affected CDC25 interaction. Four highly selective mutations were discovered, and they affected the surface-located amino acid residues 62, 63, 67, and 69. Two lines of evidence suggested that these residues may be involved in binding to CDC25: (i) using the yeast two-hybrid system, we demonstrated that these mutants cannot bind CDC25 under conditions where the wild-type H-Ras protein can; (ii) we demonstrated that the binding to H-Ras of monoclonal antibody Y13-259, whose epitope has been mapped to residues 63, 65, 66, 67, 70, and 73, is blocked by the mouse sos1 and yeast CDC25 gene products. We also present evidence that the mechanism by which CDC25 catalyzes exchange is more involved than simply catalyzing the release of bound nucleotide and passively allowing nucleotides to rebind. Most critically, a complex of Ras and CDC25 protein, unlike free Fas protein, possesses significantly greater affinity for GTP than for GDP. Furthermore, the Ras CDC25 complex is more readily dissociated into free subunits by GTP than it is by GDP. Both of these results suggest a function for CDC25 in promoting the selective exchange of GTP for GDP. 相似文献
39.
The entire mitochondrial genome of Schizosaccharomyces pombe ura4-294h
-was analyzed by the 2D pulsed field gel electrophoresis technique developed by Brewer and Fangman. The genome consists of multimers with an average size of 100 kb and analysis of the overlapping restriction fragments of the complete mitochondrial DNA (mtDNA) genome resulted in simply Y 2D gel patterns. Large single-stranded DNA molecules or double-stranded DNA molecules containing large or numerous single-stranded regions were found in the S. pombe mtDNA preparation. The replication of mtDNA monomers was found to occur in either direction. On the basis of these results, a replication mechanism for S. pombe mtDNA that is most consistent with a rolling circle model is suggested. 相似文献
40.
Factors affecting viability of IVF-derived bovine blastocysts after freezing and thawing were investigated. A total of 1,101 ova matured and fertilized in vitro were cultured under 2 different conditions, 1) in TCM-199 on granulosa cell monolayers at 5% CO(2) in air and 2) in synthetic oviduct fluid (SOF) medium without somatic cell support at 5% CO(2), 5% O(2), 90% N(2). All blastocysts that developed from the 2 different culture systems were individually classified into 4 grades of embryo quality and were then frozen by conventional slow freezing. Developmental rates of the IVF-derived ova to blastocysts and the survival rates of the frozen-thawed blastocysts were not different between the SOF medium (16 and 49%) and the co-culture system (13 and 61%, respectively). Survival of frozen-thawed blastocysts was affected by embryo quality in both the SOF and co-culture systems (P<0.001). Blastocysts produced in vitro were also individually classified into 3 developmental stages and were then cultured for 3 d in the co-culture system with granulosa cells after freezing and thawing. There was a difference in the survival rate of frozen-thawed embryos between blastocyst developmental stages (early vs mid, P<0.05; mid vs expanded, P<0.01; early vs expanded, P<0.001). The post-thawing survival rate of blastocysts frozen at Day 7 (62%) of culture was higher compared with that of Day 8 (45%), but there was no difference in survival rate between Day 7 and 8 of culture. The results indicate that the quality and developmental stage of blastocysts are important factors influencing their survival after freezing and thawing. 相似文献