禾本科植物的组织培养研究及其应用

禾本科植物是粮食作物的主要来源,随着人口的增加和生活水准的提高,人类对粮食的产量、种类和质量的需求也日益迫切。根据国外在1982年对90个发展中国家的统计和预测的结果说明到1990年末,这些国家共缺少72百万吨谷物而到2000年将缺少144百万吨谷物。近十余年以来,随着植物分子生物学的迅猛发展和作为植物生物技术重要组成部分的植物组织培养技术的日臻完善,被公认为非常困难从事的禾本科植物(Gramina-ceae)的组织培养也取得了异常迅速的发展,并且已经在作物改良的生产中取得了成效,显示了越来越大的潜能和威力,为人类从根本上解决食物问题指出了一条切实可行的途径。本文拟在评述近年来禾本利植物组织培养(主要指胚胎培养、器官培养、细胞培养和原生质体培养)的理论性研究和应用性研究的进展,并重点描述和讨沦在应用上较为成熟和有发展前景的几个领域的发展现状以及利用禾本科植物的组织培养技术而进行的基因转移技术的概况。希望能为我国从事植物组织培养的工作者们提供某些参考资料并对于一些问题进行共同的商榷和探讨。     

人α1/158Ⅴ型(α1c型)干扰素基因在大肠杆菌中的表达及其产物的初步纯化

采用DNA聚合酶链反应(PCR)技术,将本实验室从中国人胎肝细胞染色体DNA中发现和分离的IFN-α1/158V基因的原始克隆,改造成适于进行非融合蛋白原核表达的结构形式,并在大肠杆菌中获得高效表达。测得重组IFN-α1/158V的抗病毒活性为1.9×10~7单位/升菌液。随后又采用以单克隆抗体亲和层析为主的纯化流程对表达产物进行初步纯化,获得了在SDS-聚丙烯酰胺凝胶电泳上呈现单一条带的纯化产物。     

改变乙型肝炎病毒核心抗原基因3′端序列对其在原核细胞中表达的影响

乙型肝炎病毒(HBV)的核心抗原基因(C基因)编码185个氨基酸残基,在原核细胞或痘苗病毒系统中能表达并装配成27nm大小的核心抗原(HBcAg)多聚体颗粒。已证实HBV C基因3′端编码近40个氨基酸的碱基序列,不是表达形成HBcAg颗粒所必需的。用外源基因替换这部分序列,已表达出表面带有外源基因产物的杂合颗粒,它具有很好的免疫原性,成为新型的基因工程多决定簇颗粒载体疫苗。但我们的实验中发现,用另外的外源基因替换3′端序列能显著影响HBV C基因在大肠杆菌中的表达,不同组成的外源基因其影响程度有所不同。     

肾上腺皮质激素对心房钠尿肽基因表达的促进作用

给完整的及切除肾上腺的雌性 Wistar 大鼠分別注射地塞米松、去氧皮质酮或地塞米松加去氧皮质酮;冷酚法提取心房总 RNA,用α-~(32P)标记的大鼠心房肽 cDNA 探针与之杂交。完整大鼠接受地塞米松和切除肾上腺后接受地塞米松加去氧皮质酮的大鼠,心房肽基因转录产物增加2倍,其余组无显著变化。结果提示糖皮质激素可促进心房肽基因表达,但此作用依赖于盐皮质激素的同时存在,单纯盐皮质激素不能增强该基因的表达。     

Homosexual male pairing in Schistosoma mansoni

and 1988. Homosexual male pairing in Schistosoma mansoni.International Journal for Parasitology 18: 1115–1117. To see whether male worms within the gynecophoral canal of another male worm would become feminized (i.e. express vestigial female-associated genes), we established homosexual pairs by twice exposing mice to male cercariae with a 4 or 6-week interval, and perfusing 3–5 weeks later. From 13 to 34% of these worms were found in pairs, compared with 0 to 7% in singly exposed controls. ‘Inner’ males in homosexual pairs showed no histological evidence of female reproductive structures, but were stunted, had poorly developed testes, and the high nuclear density characteristic of mature females. More vitelline follicles occurred in unpaired unisexual males than in homosexually paired males, fewest in bisexually paired males. Uptake of tyrosine, an indicator of vitelline development, occurred in the same relative order. The gynecophoral microenvironment often led to stunting, probably through starvation induced by the relative inaccessibility of host blood to homosexually clasped males.     

The equine protease inhibitory system (Pi): Abnormal expressions of PiF,PiL, and PiS

Three cases of abnormal expression of the equine protease inhibitory alleles, Pi F, L, and S₁, were observed following the examination of 30,000 plasma samples by one-dimensional acid (pH 4.6) polyacrylamide gel electrophoresis. Characterization of the abnormal proteins in terms of isoelectric point, molecular mass, inhibitory spectra, and sialic acid content was performed using one- and two-dimensional electrophoretic techniques. The Pi F and S₁ abnormalities were postulated to be the result of amino acid substitutions causing alterations in the processing of the carbohydrate side chains. No explanation could be offered for the Pi L abnormality other than a charge shift mutation. Abnormal types, F*, L*, and S*₁ behaved as alleles but the distribution of L* in offspring from one stallion (present in only 6 of 83 offspring) differed significantly from expectation.This work was supported by a grant from the Australian Stud Book, Alison Road, Randwick, N.S.W. 2031.     

Cloning of Candida pelliculosa beta-glucosidase gene and its expression in Saccharomyces cerevisiae

Summary Candida pelliculosa var. acetaetherius is a strain of yeast which can utilize cellobiose as the carbon source. From a gene library prepared from this yeast, the -glucosidase gene has been cloned in a S. cerevisiae host using a chromogenic substrate, 5-bromo-4-chloro-3-indolyl--glucoside as an indicator. It was proved by Southern analysis that the DNA fragment carrying the -glucosidase gene originated from C. pelliculosa. -Glucosidase produced by S. cerevisiae transformants was secreted into the periplasmic space. In Candida, -glucosidase was not induced by cellobiose but was derepressed by lowering the concentration of glucose. The regulation of -glucosidase synthesis in S. cerevisiae carrying the cloned -glucosidase was not clear compared with that in Candida, however, the enzyme activity in low glucose medium (0.05%) was reproducibly higher than in high glucose medium (2%). We have found the sequence that controls the expression of the -glucosidase gene negatively in S. cerevisiae.     

烟草愈伤组织器官发生过程中外源激素的作用

近年来,利用愈伤组织系统在与器官发生有关的形态学、生理学和生物化学研究方面已经取得了一些进展(Thorpe 1980,刘涤 1983)。对烟草愈伤组织系统的研究明确了外源激素对器官发生类型的调节作用(Skoog 1971,Engelke等1973,刘涤等1980)。但是,这些研究仅考虑到外源激素的作用,而对器官发生过程中的其它变化并不了解。最近,Kamada和Harada(1981)比较了胡萝卜体细胞胚发育过程中内源IAA和ABA含量的变化,Noma等(1982)证实形成胚的和不形成胚的细胞间GA_3含