The carboxy terminus of pp60c-src is a regulatory domain and is involved in complex formation with the middle-T antigen of polyomavirus.

A large number of mutations were introduced into the carboxy-terminal domain of pp60c-src. The level of phosphorylation on Tyr-416 and Tyr-527, the transforming activity (as measured by focus formation on NIH 3T3 cells), kinase activity, and the ability of the mutant pp60c-src to associate with the middle-T antigen of polyomavirus were examined. The results indicate that Tyr-527 is a major carboxy-terminal element responsible for regulating pp60c-src in vivo. A good but not perfect correlation exists between lack of phosphorylation at Tyr-527 and increased phosphorylation at Tyr-416, between elevated phosphorylation on Tyr-416 and activated kinase activity, and between activated kinase activity and transforming activity. Phosphorylation of Tyr-527 was insensitive to the mutation of adjacent residues, indicating that the primary sequence only has a minor role in recognition by kinases or phosphatases which regulate it in vivo. Three mutants which have in common a modified Glu-524 residue were phosphorylated on Tyr-416 and Tyr-527 and were weakly transforming. This suggests that other mechanisms besides complete dephosphorylation of Tyr-527 can lead to increased phosphorylation of Tyr-416 and activation of the transforming activity of pp60c-src. Furthermore, the residues between Asp-518 and Pro-525 were required to form a stable complex with middle-T antigen. The proximity of these sequences to Tyr-527 suggests a model in which middle-T activates pp60c-src by binding directly to this region of the molecular and thereby preventing phosphorylation of Tyr-527. Alternatively, middle-T binding may mediate a conformational change in this region, which in turn induces an alteration in the level of phosphorylation at Tyr-527 and Tyr-416.     

Identification of genes required for cytoplasmic localization in early C. elegans embryos

We have isolated and analyzed eight strict maternal effect mutations identifying four genes, par-1, par-2, par-3, and par-4, required for cytoplasmic localization in early embryos of the nematode C. elegans. Mutations in these genes lead to defects in cleavage patterns, timing of cleavages, and localization of germ line-specific P granules. Four mutations in par-1 and par-4 are fully expressed maternal effect lethal mutations; all embryos from mothers homozygous for these mutations arrest as amorphous masses of differentiated cells but are specifically lacking intestinal cells. Four mutations in par-2, par-3, and par-4 are incompletely expressed maternal effect lethal mutations and are also grandchildless; some embryos from homozygous mothers survive and grow to become infertile adults due to absence of functional germ cells. We propose that all of these defects result from the failure of a maternally encoded system for intracellular localization in early embryos.     

Microtubule and microfilament distribution and tubulin content in the cell cycle of Indian muntjac cells

Using DAPI, rabbit antitubulin antibody, FITC-labeled goat anti-rabbit IgG, and TRITC-phalloidin to stain individual cells, the microspectrophotometric analysis showed that three markers that represent the nucleus, microtubules (MT), and microfilaments (MF), respectively, could be recognized in individual cells without interference. The phase of the cell cycle was determined by DNA content. We found that in Indian muntjac (IM) cells, the amount of tubulin in G2 and M phases was about twice as much as that in G1 phase. In G2 cells, the cytoplasmic microtubule complex (CMTC) became denser than in G1 cells. The cytoplasmic MT extent in basically the same orientation as MF bundles in interphase. The regions where the MT is denser also have a denser MF distribution.     

Laser-transected microtubules exhibit individuality of regrowth, however most free new ends of the microtubules are stable

To study the possible mechanism of microtubule turnover in interphase cells, we have used the 266-nm wavelength of a short-pulsed Nd/YAG laser to transect microtubules in situ in PtK2 cells at predefined regions. The regrowth and shrinkage of the transected microtubules have been examined by staining the treated cells with antitubulin mAb at various time points after laser irradiation. The results demonstrate that microtubules grow back into the transected zones individually; neither simultaneous growth nor shrinkage of all microtubules has been observed. The half-time of replacement of laser-dissociated microtubules is observed to be approximately 10 min. On the other hand, exposure of the core of the microtubule, which is expected to consist almost completely of GDP-tubulin, by transecting the internal regions of the microtubule does not render the remaining polymer catastrophically disassembled, and most transected microtubules with free minus ends do not quickly disappear. Taken together, these results suggest that most microtubules in cultured interphase cells exhibit some properties of dynamic instability (individual regrowth or shrinkage); however, other factors in addition to the hydrolysis of GTP-tubulin need to be involved in modulating the dynamics and the stability of these cytoplasmic microtubules.     

马铃薯未传粉子房离体培养诱导双单倍体植株

以MS为基本培养基,附加不同水平的生长素,培养未传粉马铃薯子房,经三年试验,从两个品种中获得了双单倍体的绿色小植株,(2n=2x=24);从分化绿苗力很强的球状愈伤组织中,又不断地分化出许多绿色小植株。绝大多数品种,都可以诱导出愈伤组织,一般诱导率为70％左右。品种的基因型和培养基中的生长素种类与水平在愈伤组织分化绿苗中起着重要作用。通过扦插和试管微型薯培养,可以大量繁殖试管苗,这为马铃薯单倍体育种提供了较有利的条件。     

The rabbit alpha-like globin gene cluster is polymorphic both in the sizes of BamHI fragments and in the numbers of duplicated sets of genes

The alpha-like globin gene cluster in rabbits contains embryonic zeta- globin genes, an adult alpha-globin gene, and theta-globin genes of undetermined function. The basic arrangement of genes, deduced from analysis of cloned DNA fragments, is 5'-zeta 0-zeta 1-alpha 1-theta 1- zeta 2-zeta 3-theta 2-3'. However, the pattern of restriction fragments containing zeta- and theta-globin genes varies among individual rabbits. Analysis of BamHI fragments of genomic DNA from 24 New Zealand white rabbits revealed eight different patterns of fragments containing zeta-globin genes. The large BamHI fragments containing genes zeta 0 and zeta 1 are polymorphic in length, whereas a 1.9-kb fragment containing the zeta 2 gene and the 3.5-kb fragment containing the zeta 3 gene do not vary in size. In contrast to this constancy in the size of the restriction fragments, the copy number of the zeta 2 and zeta 3 genes does vary among different rabbits. No length polymorphism was detected in the BamHI fragments containing the theta-globin genes, but again the copy number varies for restriction fragments containing the theta 2 gene. The alpha 1- and theta 1-globin genes are located in a nonpolymorphic 7.2-kb BamHI fragment. The combined data from hybridization with both zeta and theta probes shows that the BamHI cleavage pattern does not vary within the region 5'-alpha 1-theta 1- zeta 2-zeta 3-theta 2-3', but the pattern genomic blot-hybridization patterns for the progeny of parental rabbits with different zeta-globin gene patterns shows that the polymorphic patterns are inherited in a Mendelian fashion. Two different haplotypes have been mapped based on the genomic blot-hybridization data. The variation in the alpha-like globin gene cluster in the rabbit population results both from differences in the copy number of the duplication block containing the zeta-zeta-theta gene set and from the presence or absence of polymorphic BamHI sites.      

油菜花序轴、叶片组织培养中形态发生的细胞组织学研究

油菜花序轴、叶片在MS培养基上经诱导可产生愈伤组织并形成再生植株。愈伤组织形成可分3个时期:细胞启动期、分裂期和分化期。实验证明:花序轴启动部位多发生在皮层薄壁细胞、髓部薄壁细胞、韧皮薄壁细胞和木薄壁细胞。叶片主要是叶肉细胞、叶脉薄壁细胞和表皮细胞启动。细胞分裂期的特点是启动细胞反复增殖,形成连续的分生细胞层。细胞分化期的特点是一部分细胞保持分裂能力,一部分细胞分化形成薄壁细胞和输导分子。观察了愈伤组织中器官发生的部位和方式。实验证明,根的发生多为内起源,芽的发生多为外起源。     

用体外免疫法建立人—人抗流行性乙型脑炎病毒杂交瘤细胞株

利用单克隆抗体(McAb)进行病毒病的治疗是人们所关心的一个重大课题。 流行性乙型脑炎(乙脑)是一种严重威胁人民健康的急性传染病,病死率高,后遗症严重。国内外目前尚无特效疗法。陈伯权等用乙脑病毒皮下或腹腔感染3周龄小白鼠24、48小时及5天后,分别用乙脑病毒51-8McAb进行治疗,平均治愈率分别为78％、73％及22％。     

中水技术污水处理工艺去除病毒的效果

中水技术开发既可缓解水资源的紧缺,又能改善城市环境。本文报道用石英粉吸附/洗脱和浓缩棒脱水的二步浓集水病毒技术(回收率为25.4～37.3％),研究中水技术不同处理工艺去除病毒的效果。结果,二级处理可去除掺入病毒的98.9％,结合三级处理的混凝沉淀和过滤,可去除99.976％,再加活性炭吸附或加氯消毒,所得中水分别去除掺入病毒的99.986％和99.991％。经首都机场中水道试验厂水样检测表明,中水中未检测到病毒(<0.23PFU/L)。