Primary structure of an EcoRI fragment of lambda imm434 DNA containing regions cI-cro of phage 434 and cII-o of phage lambda.

Digestion of phage lambda imm434 DNA with restriction endonuclease EcoRI yields 7 fragments. The shortest among them (1287 bp) contains the right part of the phage 434 immunity region and the phage DNA portion proximal to it. The complete primary structure of this fragment has been determined using the chemical method of DNA sequencing. Hypothetical amino-acid sequences of proteins coded by the cro gene of phage 434 and the cII gene of phage lambda, as well as NH2-terminal amino-acid sequences of the cI protein of phage 434 and the O protein of phage lambda, have been deduced solely on the basis of the DNA sequence. The fragment studied contains also the pR and probably prm promoters and the oR operator of phage 434. The sequence coding for them differs from the respective DNA sequence of phage lambda.     

cDNA approaches to isolation of the mouse mutant weaver gene

The mouse autosomal recessive mutant gene weaver (wv) results in abnormalities in cerebellum, substantia nigra and testis. Although a subtracted cDNA library prepared by removing P31 (wv/wv) sequences from a P1 (wv/+) library should contain mainly nonrepetitive neonatal sequences, unfortunately, repetitive sequences still appear during screening. Two clones, one repetitive, the other not, are used to illustrate the problems encountered in attempting to isolate the weaver gene from a subtracted cDNA library.Special issue dedicated to Dr. Sidney Ochs.     

分离自安徽土壤金龟子绿僵菌种群的遗传异质性研究

金龟子绿僵菌是昆虫种群自然控制和害虫生物防治中重要的虫生真菌,已被作为化学农药的替代品广泛应用于农林害虫的防治。其寄主范围和地理分布极广,因此种群结构也十分复杂。为明确安徽省土栖金龟子绿僵菌的种群遗传结构,采用ISSR分子标记技术对采集自安徽省不同地区土壤中的116株金龟子绿僵菌进行遗传异质性分析。分子数据显示,筛选出的8个引物共获得79个位点,其中多态位点比率为100%。不同地区种群的Nei’s基因多样性(H)为0.2796,Shannon信息指数(I)为0.4425,种群间存在一定程度的基因流(Nm=4.4282)和遗传分化(Gst=0.1015),种群内的基因多样度占总居群的89.85%,种群间占10.15%,表明安徽土栖金龟子绿僵菌的遗传变异主要来源于种群内,且种群内表现出较大的遗传变异。并采用UPGMA对所有供试材料进行聚类分析,得到7大类种群。结果表明,安徽土栖金龟子绿僵菌之间的亲缘关系与地理来源不存在相关性。     

常压低氧小鼠氧耗量测定新装置的研究及应用

目的：建立一种实时记录常压低氧环境中动物氧耗量的方法。方法：本实验装置由动物舱、补水控制系统、天平、软管、装有体重记录软件的电脑等组成。为了实现常压低氧,用水补充动物消耗的氧气以保持动物舱内压力恒定,这个过程由气液联动装置控制;补充的水量由天平测量并同步输出信号至excel文档中。用注射器抽气校准方法检测了装置的准确性和精度。利用该装置观察了6只急性重复低氧小鼠（处理组）和6只未经低氧处理的小鼠（对照组）的常压低氧过程的氧耗量特征。结果：不同体积抽气量与相应补水量两组数据配对t检验P=1;重复抽1 ml氧气6次的补水量变异系数为4%。处理组小鼠的存活时间为（58.8±6.8）min,显著高于对照组（46.0±8.7）min（P〈0.05）。处理组小鼠的总氧耗量为（85.1±8.5）ml,显著高于对照组（73.6±5.4）ml（P〈0.05）。结论：处理组小鼠摄取氧总量增多从而显著延长其存活时间。氧耗量测定装置准确度和精密度较高,可用于低氧研究中氧耗量的测定。     

结合SSR标记和STS标记对家蚕无鳞毛翅基因的定位

家蚕突变表型无鳞毛翅(non-lepis wing, nlw)由隐性基因nlw控制。由于家蚕雌性不发生交换, 文章采用有鳞毛翅品系P50和无鳞毛翅品系U06两个品系组配F1代及BC₁回交群体, (U06×P50)×U06和U06×(U06×P50)分别记作BC₁F和BC₁M, 根据已经构建的家蚕SSR分子标记连锁图谱及已经发表的有关序列对nlw基因进行了连锁及定位分析。得到8个与nlw基因连锁的SSR(Simple sequence repeat)标记和1个STS(Sequence-tagged sites)标记。BC₁F群中的所有正常翅个体均表现出与(U06×P50)F₁相同的杂合带型; 而所有无鳞毛个体带型与亲本U06一致, 为纯合型。利用BC₁M群体构建了关于nlw基因的遗传连锁图, 连锁图的遗传距离为125.7 cM, 与nlw基因最近的引物为STS标记cash2p, 图距为11.4 cM。     

Chitinase III in pomegranate seeds (Punica granatum Linn.): a high-capacity calcium-binding protein in amyloplasts

Chitinases are a class of ubiquitous proteins that are widely distributed in plants. Defense is the major natural role for chitinases, primarily against fungal pathogens. Little is known regarding their non-defensive roles in seeds. In this study, a new class III chitinase from pomegranate seeds (pomegranate seed chitinase, PSC) was isolated and purified to homogeneity. The native state of PSC is a monomer with a molecular weight of approximately 30 kDa. This chitinase naturally binds calcium ions with high capacity and low affinity, suggesting that PSC is a calcium storage protein. Consistent with this idea, its amino acid sequence (inferred from cDNA) is rich in acidic amino acid residues, especially Asp, similar to reported calcium storage proteins. The presence of calcium considerably improves the stability of the protein but has little effect on its enzymatic activity. Transmission electron microscopy analyses indicate that, similar to phytoferritin, this enzyme is widely distributed in the stroma of amyloplasts of the embryonic cells, suggesting that amyloplasts in seeds could serve as an alternative plastid for calcium storage. Indeed, the transmission electron microscopy results showed that, within the embryonic cells, calcium ions are mainly distributed in the stroma of the amyloplasts, consistent with a role for PSC in calcium storage. Thus, the plant appears to have evolved a new plastid for calcium storage in seeds. During seed germination, the content of this enzyme decreases with time, suggesting that it is involved in the germination process.     

An evaluation of the utility of SSR loci as molecular markers in maize (Zea mays L.): comparisons with data from RFLPS and pedigree

 The utility of 131 simple sequence repeat (SSR) loci to characterize and identify maize inbred lines, validate pedigree, and show associations among inbred lines was evaluated using a set of 58 inbred lines and four hybrids. Thirteen sets of inbred parent-progeny triplet pedigrees together with four hybrids and their parental lines were used to quantify incidences of scoring that departed from expectations based upon simple Mendelian inheritance. Results were compared to those obtained using 80 restriction fragment length polymorphism (RFLP) probes. Over all inbred triplets, 2.2% of SSRs and 3.6% of RFLP loci resulted in profiles that were scored as having segregated in a non-Mendelian fashion. Polymorphic index content (PIC, a measure of discrimination ability) values ranged from 0.06 to 0.91 for SSRs and from 0.10 to 0.84 for RFLPs. Mean values for PIC for SSRs and RFLPs were similar, approximately 0.62. However, PIC values for nine SSRs exceeded the maximum PIC for RFLPs. Di-repeats gave the highest mean PIC scores for SSRs but this class of repeats can result in “stutter” bands that complicate accurate genotyping. Associations among inbreds were similar for SSR and RFLP data, closely approximating expectations from known pedigrees. SSR technology presents the potential advantages of reliability, reproducibility, discrimination, standardization and cost effectiveness over RFLPs. SSR profiles can be readily interpreted in terms of alleles at mapped loci across a broad range of maize germ plasm. Consequently, SSRs represent the optimum approach for the identification and pedigree validation of maize genotypes compared to other currently available methods. Received: 15 January 1997 / Accepted: 28 February 1997     

Sequence-tagged-site (STS) markers of arbitrary genes: the utility of black spruce-derived STS primers in other conifers

 Sequence-tagged-site primers, previously developed based upon black spruce (Picea mariana) cDNA sequences, were tested for their ability to direct specific amplification in two individuals of each of 12 additional conifer species. Nearly all (95–97%) of the primers functioned well in congeneric trials, while a lower proportion (21–33%) scored positively in other Pinaceae genera. Outside of the Pinaceae, amplification of homologous products was not achieved. Products from the various species often differed in size from their homologs in black spruce. In one case a large difference in size was due to the lack of an intron in a jack pine product while in several other cases the differences were due to the presence or absence of large direct repeats in the DNA sequences. Length polymorphism was occasionally evident between the two individuals examined of a given species. We investigated marker polymorphism in detail in a panel of 15 white spruce (Picea glauca) trees. Allelic segregation among haploid megagametophytes was revealed directly at 16 loci by standard agarose-gel electrophoresis without any additional manipulation of amplification products. Polymorphisms observed at 12 of these loci were exclusively co-dominant. For this subset of 12 loci, the average number of alleles was 3.2 and the average observed heterozygosity was 0.37. Received: 10 April 1998 / Accepted: 22 April 1998