精子介导GFP基因在小鼠早期胚胎中的表达

利用 DIG 末端标记技术和免疫组化技术分析了小鼠精子体外结合内化外源DNA的效率。试验结果表明,不同小鼠个体的精子结合外源DNA的阳性率有明显差异（P<0.01）,平均为13%。利用考马斯亮蓝染色评价了小鼠精子顶体反应发生的情况,筛选出TYH培养液为较合适的体外受精液。利用小鼠体外受精技术,将体外转染GFP基因并获能的小鼠精子与成熟卵母细胞进行体外受精,受精卵进行体外培养,表达GFP胎的阳性率为4.7%。验证了精子介导制备转基因小鼠胚胎的可行性,并建立了利用精子载体法制备转基因小鼠胚胎的平台。     

Asymmetric Somatic Hybridization Between Wheat (Triticum aestivum) and Bromus inermis

Asymmetric hybridization was conducted between wheat and Brorrats inermis keyss which is a distanfiy related intergeneric plant (belonging to different tribe) of wheat and possesses some favorable traits, such as resistant to cold, drought and disease. Protoplasts isolated from young embryo-derived calli of common wheat ( Triticura aestivum L., tv. 99P, (AABBDD), 2n = 42) were fused with UV-treated protoplasts isolated from young embryo-derived calli of Bromus inermis by PEG method. Three clones (No. 1 ~ No. 3) were regenerated from the fusion products and differentiated into albino seedlings. The clones and the seedlings were all verified as hybrids by chromosome counting, isozyme and RAPD analysis. Their isozyme and RAPD pattern contained the characteristic bands of both parents as well as new band(s). The chromosome numbers of albino were in the range of 42~54 with small chromosomes of Bromus inerm/s and chromosome fragments. The above results confirmed that hybrid albinos were obtained.     

Assembly characteristics of plant keratin intermediate filaments in vitro

After selective extraction and purification, plant keratin intermediate filaments were reassembled in vitro. Scanning tunneling microscope (STM) and transmission electron microscope (TEM) micrographs showed that acidic keratins and basic keratins can assemble into dimers and further into 10 nm filaments in vitro. In higher magnification images, it can be seen that fully assembled plant keratin intermediate filaments consist of several thinner filaments of 3 nm in diameter, which indicates the formation of protofilaments in the assembly processes. One of the explicit features of plant keratin intermediate filaments is a 24—25 nm periodic structural repeat alone the axis of beth the 10 nm filaments and protofilaments. The periodic repeat is one of the fundamental characteristic of all intermediate filaments, and demonstrates the half staggered arrangement of keratin molecules within the filaments.     

Amplification, analysis and chromosome mapping of novel homeobox-containing and homeobox-flanking sequences in rice

Homeobox genes, widely distributed among animal and plant kingdoms, play an important role in developmental process. Several homeobox conserved fragments were amplified by PCR and the flanking regions were also obtained by an LM-PCR procedure. Sequencing and Southern analysis showed that they belong to a homeobox gene family of rice. Six homeobox-containing fragments were mapped on the molecular linkage map of rice. They were located on chromosomes 3, 4 and 7 respectively. It is noteworthy that there are 4 homeobox fragments located on rice chromosome 3 and the result is also consistent with the comparative genomics between rice and maize.     

Composition and structure of nucleolar skeleton (nucleolar matrix)——Actin and fibrillarin are two main protein components of nucleolar skeleton

Purified nucleoli of HeLa cells were treated sequentially with nonionic detergent, nucleic acid enzyme, low salt and high salt. The residual nucleolar structure termed nucleolar skeleton (nucleolar matrix) was shown as a fine network under electron microscope with DGD embedding-unembedding technique. Such structures of BHK-21 cell and mouse liver cell are similar to that of HeLa cell. The protein composition of the nucleolar skeleton of HeLa cells was analyzed. The protein composition of such nucleolar residual shows obvious difference from the compositions of nuclear matrix and chromosome scaffold. The major protein composition of the nucleolar skeleton of HeLa cells contains 6-7 polypeptides. Their molecular weights are about 48, 43, 36 and 33 ku. Further studies show that actin and fib-rillarin are two major protein components of nucleolar skeleton of HeLa cells.     

水稻抗白叶枯病基因Xa-4的PCR标记研究

根据与水稻抗白叶枯病基因Xa-4紧密连锁的分子标记M55的序列设计引物,通过对国际水稻研究所育成的抗白叶枯病近等基因系和基因累加系的叶片DNA、半粒种子提取物及Xa-4基因的杂合体DNA的PCR特异扩增,初步建立了Xa-4的PCR标记体系。进而用该标记体系对我国籼型杂交水稻常用的亲本材料进行分析,揭示出了Xa-4在这些材料中的分布情况。 Abstract　Based on the sequence of a DNA marker tightly linked to the rice bacterial blight（BB） resistance gene Xa-4, two primers were designated and synthesized to develop a PCR marker for the gene. Specific amplified polymorphism analysis was carried out with these primers on a set of BB resistance isogenic lines and pyramided lines developed by IRRI. Two PCR bands were revealed corresponding to lines with dominant Xa-4 and those with the recessive allele, respectively, regardless the lines pyramided with other resistance genes. A hybrid with heterozygous Xa-4 produced both of the two allele PCR pattern. Then, the PCR marker was used to survey a range of hybrid rice germplasm. The results of the germplasm survey will be useful in hybrid rice breeding programs aimed at exploiting Xa-4.     

拟南芥细胞中存在中间纤维的研究

利用整装电镜制样与选择性抽提技术,在拟南芥（Ａｒａｂｉｄｏｐｓｉｓｔｈａｌｉａｎａ （Ｌ．） Ｈｅｙｎｈ） 愈伤组织细胞质中观察到直径１０ ｎｍ 左右的纤维网络结构。免疫印迹分析表明纤维的主要成分是６ 种多肽,它们分别与动物角蛋白单克隆抗体ＡＥ１ 、ＡＥ３ 有免疫交叉反应。利用间接免疫荧光技术,与ＡＥ１ 和ＡＥ３ 反应的抗原呈弥散状定位于整个细胞质中,而且１０ ｎｍ 纤维可以在体外重新组装。以上结果表明,在拟南芥细胞质中存在类角蛋白的中间纤维。以动物中间纤维基因的保守序列为引物,采用ＲＴ＿ＰＣＲ技术,进一步从这一模式植物中克隆到一个ｃＤＮＡ片段,这可能为从分子水平上证明植物中间纤维的存在提供了一个线索     

单倍体小麦与羊草的属间体细胞杂交

以失去植株再生能力的小麦单倍体愈伤组织和羊草二倍体愈伤组织为材料,游离原生质体,并用紫外线自理２羊草原生质体,用ＰＥＧ法融合,对融合克隆进行染色体和同工酶分析,在已贩２６个克隆中有２１个是杂种,其中有一个克隆再生出短命小植株,结果 体小麦与二倍体草的不对称融合虽然再生互补效应不如二倍体小麦,然而杂种优先生长的现象仍然比较明显。如果改善实验条件和双亲原始的再生能力,这种融合方式仍然可以利用。     

盐度对互花米草枯落物分解释放硅、碳、氮元素的影响

为了研究潮汐湿地盐度对枯落物分解过程中硅、碳、氮元素释放的影响,通过室内模拟不同盐度(0、5、15和30)对互花米草枯落物茎和叶分解释放过程中硅、碳、氮元素的动态变化进行测定。结果表明:(1)互花米草茎和叶枯落物失重率和分解速率均随盐度增加而降低。(2)互花米草茎和叶枯落物分解水体中硅含量均随着盐度升高而增加,并且盐度30处理下,枯落物分解硅释放量显著高于盐度0和5(P0.05)。而分解末期生物硅残留量则随盐度升高而降低。(3)不同盐度处理茎枯落物分解碳释放量无显著差异,但叶枯落物分解碳释放量在盐度5、15和30处理中显著高于淡水(P0.05)。(4)互花米草茎枯落物分解释放到水中的NH_4~+-N含量随着盐度的升高而减少,NO_3~--N含量与之相反。研究单因素盐度对枯落物分解及元素释放的影响,可以为预测潮汐湿地枯落物分解对盐水入侵的响应机制提供参考,为湿地生源要素生物地球化学循环过程研究提供基础依据。