Oligodeoxynucleotide-directed cleavage and repair of a single-stranded vector: a method of site-specific mutagenesis

A simple and efficient site-specific mutagenesis method is described. First, a single-stranded (ss) circular vector is linearized at the site where the desired mutation will be introduced. To do this, an oligodeoxynucleotide complementary to the target region of the ss vector and containing a restriction enzyme recognition sequence is annealed to the circular ss vector, and the partial double-strand formed is subsequently cleaved with that enzyme. Then, another oligodeoxynucleotide spanning the nick and carrying the mutation is annealed to the linearized ss DNA template and the annealed mixture is used directly to transform Escherichia coli without prior enzymatic DNA synthesis in vitro. The procedure has been applied successfully to constructing insertion, deletion, and point mutations in both M13 phage vectors and plasmid vectors containing the f1 origin of replication.     

The macrosatellites of the Toulouse goose: the major tandemly repetitive DNA in the toulouse goose genome

This paper examines the principal classes of repetitive DNA of the Toulouse goose (Anser anser) genome. There are four major classes and they are tandem repeats of less than 200 base pairs (bp). The longest repeat (class A) is 190 bp long and starts with a HinfI site. Class B is 43 bp long, commencing with a FokI site. Classes A and B show no extensive homology to DNA sequences held on a current data base (Genbank) but were confirmed to exist as major repeats in another strain of goose, the Emden goose (Anser anser) genome. Classes C and D are 5-bp repeats of 5' GAGAG 3' and 5' GGGAA 3', respectively. The macrosatellites C and D were compared with a current data base (Genbank) and were found to exist in a variety of other organisms as satellites.     

Close association of a DNA replication origin and an ARS element on chromosome III of the yeast, Saccharomyces cerevisiae.

Two dimensional gel electrophoretic techniques were used to locate all functional DNA replication origins in a 22.5 kb stretch of yeast chromosome III. Only one origin was detected, and that origin is located within several hundred bp of an ARS element.     

On the generalization of the Weinbaum-Jiji bioheat equation to microvessels of unequal size; the relation between the near field and local average tissue temperatures

The extensive series of experiments reported in Lemons et al. [1] show that measureable local tissue temperature fluctuations are observed primarily in the vicinity of the 100-500 micron countercurrent vessels of the microcirculation and thus strongly support the basic hypothesis in the new bioheat equation of Weinbaum and Jiji [2] that these countercurrent microvessels are the principal determinants of local blood-tissue heat transfer. However, the detailed temperature profiles in the vicinity of these vessels indicate that large asymmetries in the local temperature field can result from the significant differences in size between the countercurrent artery and vein. Using the superposition techniques of Baish et al. [9], the paper first presents a solution to the classic problem of an unequal countercurrent heat exchanger with heat loss to the far field. This solution is then used to generalize the Weinbaum-Jiji bioheat equation and the conductivity tensor that appears in this equation to vessels of unequal size. An asymptotic analysis has also been developed to elucidate the relationship between the near field temperature of the artery-vein pair and the local average tissue temperature. This analysis is used to rigorously prove the closure approximation relating the local arterial-venous temperature difference and the mean tissue temperature gradient which had been derived in [2] using a more heuristic approach.     

从“湖北光敏感核不育水稻”的未受精子房和花药培养出单倍体植株

采用10种诱导培养基,培养湖北光敏感核不育水稻农垦58品种的未受精子房和花药。共培养未受精子房2790个,获得胚囊愈伤组织17块,最高诱导频率达3.33%,其中2块分化出绿苗。培养花药16740个,获得花药愈伤组织15块,最高诱导频率为0.92%,其中3块分化出苗,2丛白苗,1株绿苗。胚囊植株和花粉植株经根尖染色体检查为单倍体,2n=x=12。实验证明,液体培养、2,4-D0.2-0.5 mg/1、低温预处理对诱导胚囊愈伤组织及花粉愈伤组织的形成具良好效果。     

除虫菊的染色体数目及其核型

除虫菊(Pyrethrum cinerariifolium Trev.)为菊科小黄菊属多年生宿根草本植物。以花或全草入药。除虫菊的主要有效成分为除虫菊酯和瓜叶除虫菊酯,可用于加工成各种制剂作杀虫剂原料,用以防治日常虫豸(蚊、蝇、虱等)和农业害虫,且对人     

电损毁海马CA3区及连合前穹窿对大鼠血浆胰岛素水平...

Bilateral electrical lesioning of the hippocampal CA3 region (HCA3-EL) or anterior commissura hippocampi (ACHF-EL) caused marked elevations in plasma basal levels of insulin. 2 weeks later, fasting blood glucose levels were also augmented with decreased glucose tolerance. In contrast, the secretory response of pancreatic B cells to glucose stimulation was markedly enhanced. Following intravenous glucose tolerance test (IVGTT), the relative amounts of glucagon-like and insulin-like immunoreactants were reduced in the pancreatic islets of both HCA3-EL and ACHF-EL rats in comparison with the controls. In the HCA3-EL group, the relative amounts of somatostatin-like immunoreactants and gross numbers of such immunostained cells in islets were also decreased as compared with the control. No difference was seen in pancreatic-polypeptide-like immunoreactivities as assessed by immunohistochemistry plus microphotometry method. The above results suggest strongly that HCA3 and ACHF exert a tonic inhibitory action on the insulin secretion in the rat.