一组新的核仁组织者区结合蛋白

本文报道了一组新的核仁组织者区蛋白(ANOP)。这些蛋白由三种多肽组成,分子量分别为65,76和78KD。它们集中分布在核纤丝中心周围的高密度纤维组分中。研究表明ANOP广泛地分布于各种脊椎动物细胞中,有较强的抗原保守性。但在两栖类的红细胞和原肠期以前的早期胚胎细胞中则缺乏此种抗原。而在原肠形成过程中,ANOP开始出现并逐渐增加,表明ANOP可能与rRNA基因的活性有关。     

Role of viral antigens in destructive cellular immune responses to adenovirus vector-transduced cells in mouse lungs.

Adenoviruses missing E1 have been used as gene delivery vectors to the lungs for the treatment of cystic fibrosis. Transient expression of the recombinant gene and the development of inflammation have been two major limitations to the application of first-generation recombinant adenoviruses for gene therapy. Studies with mouse models of liver- and lung-directed gene therapy suggested that CD8+ cytotoxic T lymphocytes (CTLs) are effectors that contribute to extinction of transgene expression. The precise antigens responsible for activation of CTLs have not been identified. In this study, we examine the relative contributions of viral proteins versus the transgene product to the activation of CTLs which eliminate transgene-containing cells in mouse lungs. Instillation of a lacZ-expressing virus into the lungs of C57BL/6 mice elicited CTL responses to both viral proteins and the transgene product, beta-galactosidase, which collectively contribute to loss of trans-gene expression in mouse airways. Similar results were obtained in two experimental models in which the animals should be tolerant to the transgene, i.e., lacZ virus delivered to an animal transgenic for lacZ and a virus expressing the liver-specific enzyme ornithine transcarbamylase administered to the lungs of various strains of immune-competent mice. These data confirm the hypothesis that CTLs specific for viral antigens contribute to the problem of transgene instability in mouse lungs and indicate that CTLs specific for transgene product alone cannot account for the observed problem.     

In vitro antigen challenge of human antibody libraries for vaccine evaluation: the human immunodeficiency virus type 1 envelope.

Human antibody responses, or versions thereof, can be cloned as phage display libraries. In vaccine evaluation, the possibility therefore exists of challenging the human response in vitro, rather than in vivo, in order to assist in establishing the most promising vaccine leads. The characteristics of the antibodies retrieved directly indicate the strengths and weaknesses of the vaccine at the molecular level. We applied this approach to compare recombinant and native human immunodeficiency virus type 1 envelope preparations. We conclude that recombinant gp160, gp140, and, to a lesser extent, gp120 present epitopes around the CD4 binding site in a conformation different from that of the native multimer and contrary to expected vaccine requirements. Antibodies to the potently neutralizing b12 epitope were selected preferentially from an immune library by purified human immunodeficiency virus type 1 virions. This suggests that b12 is a major epitope on the virions, in contrast to recombinant envelope preparations, in which related, weakly neutralizing epitopes predominate. Although the majority of virions in the preparation used are expected to be noninfective, it appears that they predominantly express a native envelope configuration and would be able to elicit potent neutralizing antibodies.     

Patterns of viral replication correlate with outcome in simian immunodeficiency virus (SIV)-infected macaques: effect of prior immunization with a trivalent SIV vaccine in modified vaccinia virus Ankara.

The dynamics of plasma viremia were explored in a group of 12 simian immunodeficiency virus (SIV)-infected rhesus macaques (Macaca mulatta) that had received prior immunization with either nonrecombinant or trivalent (gag-pol, env) SIV-recombinant vaccinia viruses. Three distinct patterns of viral replication observed during and following primary viremia accounted for significant differences in survival times. High-level primary plasma viremia with subsequently increasing viremia was associated with rapid progression to AIDS (n = 2). A high-level primary plasma virus load with a transient decline and subsequent progressive increase in viremia in the post-acute phase of infection was associated with progression to AIDS within a year (n = 6). Low levels of primary plasma viremia followed by sustained restriction of virus replication were associated with maintenance of normal lymphocyte subsets and intact lymphoid architecture (n = 4), reminiscent of the profile observed in human immunodeficiency virus type 1-infected long-term nonprogressors. Three of four macaques that showed this pattern had been immunized with an SIV recombinant derived from the attenuated vaccinia virus, modified vaccinia virus Ankara. These data link the dynamics and extent of virus replication to disease course and suggest that sustained suppression of virus promotes long-term, asymptomatic survival of SIV-infected macaques. These findings also suggest that vaccine modulation of host immunity may have profound beneficial effects on the subsequent disease course, even if sterilizing immunity is not achieved.     

Effects of temperature, multiple allelochemicals and larval age on the performance of a specialist caterpillar

To understand the mechanisms underlying plant-insect herbivore interactions, it is necessary to examine the simultaneous effects of temperature, food quality and larval age. We examined the simultaneous effects of three allelochemicals (tomatine, rutin and chlorogenic acid) on the performance of first and second instar Manduca sexta larvae under two representative thermal regimes 21 : 10°C and 26 : 15°C for spring and summer, respectively. Thermal regime and allelochemicals interacted to influence the time from egg hatch to ecdysis to the third instar. On average, it took about half as much time to reach the third instar at 26 : 15°C as it did at 21 : 10°C. Separately, tomatine and rutin had a negative effect on developmental time from egg hatch to the third instar, but their simutaneous effects were not additive. Chlorogenic acid significantly reduced the negative effect of tomatine. The magnitude of the allelochemical effect was larger at the cooler thermal regime compared to the warmer regime. For instance, chlorogenic acid by itself had no effect at the 26 : 15°C regime, but at the 21 : 10°C regime it significantly shortened total developmental time. The effect of chlorogenic acid on stadium duration was distinctly different for the two instars. Chlorogenic acid shortened stadium duration of first instar larvae. However, depending on thermal regime and the presence of tomatine, chlorogenic acid had a negative, positive or neutral effect on stadium duration of second instar larvae. Molting duration of second instar larvae was shortened by a half day at the warmer thermal regime but was not affected by the allelochemicals. Final larval weight was influenced by rutin and chlorogenic acid. Caterpillars fed diets containing 20 moles of rutin were on average 10% lighter than those fed plain diet, whereas those fed diets containing 20 moles of chlorogenic adic were on average 7% heavier. However, the effect of chlorogenic acid depended on thermal regime. Overall, our results indicated that: 1) temperature and food quality can interact to influence insect performance and 2) these effects are influenced by larval age.     

Genomic organization of mouse Dishevelled genes

We report the isolation and characterization of two new genomic loci corresponding to the mouse Dishevelled (Dvl) genes Dvl2 and Dvl3. The Dvl genes are homologs of the Drosophila dsh segment polarity gene, and are involved in the Wnt/wingless signal transduction pathway. Dvl2 and Dvl3 genomic clones were isolated from a mouse 129 strain λFIXII genomic library and have identical exon/intron organization to Dvll. All three Dishevelled genes span 15 exons and 14 introns and have a number of conserved splice junction sites.     

大豆子叶内酸性磷酸酶活性的超微结构定位

开花后３５～５０ ｄ 期间和萌发早期（播种后４～８ ｄ）的大豆（Ｇｌｙｃｉｎｅｍ ａｘ Ｌ．）种子中,酸性磷酸酶主要分布在子叶细胞中的蛋白体内;在内质网内也检测到酸性磷酸酶活性。此外,在萌发早期的部分子叶细胞的质膜外侧及其细胞壁基质中可见密集的酸性磷酸酶活性;而且在近质膜的胞质中常见到一些富含磷酸铅沉淀的胞质小泡,似与质膜融合     

Identification of overlapping DNA-binding and centromere-targeting domains in the human kinetochore protein CENP-C.

The kinetochore in eukaryotes serves as the chromosomal site of attachment for microtubules of the mitotic spindle and directs the movements necessary for proper chromosome segregation. In mammalian cells, the kinetochore is a highly differentiated trilaminar structure situated at the surface of the centromeric heterochromatin. CENP-C is a basic, DNA-binding protein that localizes to the inner kinetochore plate, the region that abuts the heterochromatin. Microinjection experiments using antibodies specific for CENP-C have demonstrated that this protein is required for the assembly and/or stability of the kinetochore as well as for a timely transition through mitosis. From these observations, it has been suggested that CENP-C is a structural protein that is involved in the organization or the kinetochore. In this report, we wished to identify and map the functional domains of CENP-C. Analysis of CENP-C truncation mutants expressed in vivo demonstrated that CENP-C possesses an autonomous centromere-targeting domain situated at the central region of the CENP-C polypeptide. Similarly, in vitro assays revealed that a region of CENP-C with the ability to bind DNA is also located at the center of the CENP-C molecule, where it overlaps the centromere-targeting domain.     

Process conditions affecting proteolysis of recombinant proteins in Escherichia coli

The effects of process conditions on proteolysis of three recombinant IgG-binding proteins ZT3, ZZT3 and protein A in E. coli were studied. SDS/PAGE shows that the relative amount of degradation intermediates of ZT3 in a shake flask cultivation is much higher than that in a bioreactor culture. The rate of proteolysis of ZZT3 and protein A was also higher in shake flask cultures than in bioreactor cultures. The proteolysis rate constant of ZZT3 was 12/h in a shake flask but only 2.1/h in a bioreactor. Corresponding values for protein A were 2.4/h and 1.0/h, respectively. High proteolysis rate constants correlated with lower product yields.