白魔芋和花魔芋葡甘露聚糖研究

从白魔芋和花魔芋的块茎中分离纯化出两种魔芋葡甘露聚糖(Konjac Glucomannan,简称KGM),分别名为白魔芋葡甘露聚糖(aKGM)和花魔芋葡甘露聚糖(rKGM)。通过超离心、玻璃纤维纸电泳和凝胶过沪证明aKGM和rKGM都是均一的多糖。这两种多糖都由甘露糖(M)和葡萄糖(G)组成,其克分子比G/M:aKGM为1:1.69,rKGM为1:1.60,分子量前者为8.09×10~5,后者为7.37×10~5。酶水解实验和红外光谱分析说明aKGM和rKGM都是由甘露糖和葡萄糖以β-1,4-糖苷键连接的杂多糖;有O-乙酰基的特征吸收峰,说明在某些糖残基上可能有乙酰基团。     

高稳定人胰岛素的晶体学研究:Ⅲ,A21-Asp突变体的结晶和初步晶体学分析

研究一系列有明确意义改性胰岛素的结构将有益于新型胰岛素分子的设计。经蛋白质工程制备的AspA21—人胰岛素(ADHI)基本保持了生物活力,而在酸性溶夜(pH3—4)中的稳定性比天然胰岛素在中性溶液中高5—10倍。本文报道ADHI的结晶、初步晶体学研究及中等分辨率衍射数据的收集。ADHI晶体属R3空间群:a_H=b_H=82.72埃,c_H=34.02埃。     

Genomic DNA sequence for human C-reactive protein

The gene for the prototype acute phase reactant, C-reactive protein, has been isolated from two lambda phage libraries containing inserted human DNA fragments using synthetic oligonucleotide probes. Nucleotide sequence analysis indicates that after coding for a signal peptide of 18 amino acids and the first two amino acids of the mature protein, there is an intron of 278 base pairs followed by the nucleotide sequence for the remaining 204 amino acids. The intron is unusual in that it contains on the positive strand a poly(A) stretch 16 nucleotides long and a poly(GT) region 30 nucleotides long which could adopt the Z-form of DNA. The nucleotide sequence reported here confirms the amino acid sequence of mature C-reactive protein as originally reported except that it codes for an additional 19 amino acids beginning at position 62. Thus DNA sequence analysis predicts that the mature protein consists of 206 amino acids rather than 187 as originally reported. The mRNA cap site is located 104 nucleotides from the start of the signal peptide and there is a 3' noncoding region 1.2 kilobase pairs in length. The gene has a typical promoter containing the sequences TATAAAT and CAAT 29 and 81 base pairs upstream, respectively, of the cap site.     

An improved DNA sequencing strategy

A modification of Hong's systematic DNA sequencing strategy is described. The original procedure has been simplified and transfectant yield increased. After DNase I limited cleavage in the presence of Mn2+, the single-cut linear DNA does not have to be separated from supercoiled or open circular DNA on an agarose gel. After ligation, the DNA is digested with a second restriction endonuclease for which a unique cleavage site resides between the insert and the first restriction endonuclease cutting site. The original intact DNA is linearized whereas the deleted subclone is not. The background is decreased to an undetectable level. This DNA sequencing strategy was tested on a 1.4-kb DNA fragment containing the araC regulatory gene from Erwinia carotovora. A set of subclones sufficient to sequence the fragment on both strands was produced in 2 days and the yield was at least 60-fold higher than in the original protocol.     

Regeneration of the arterial wall in microporous,compliant, biodegradable vascular grafts after implantation into the rat abdominal aorta

Summary The ultrastructure of a new type of vascular graft, prepared from a mixture of polyurethane (95 weight %) and poly-L-lactic acid (5 weight %), was examined six weeks after implantation into the abdominal aorta of rats. These microporous, compliant, biodegradable, vascular grafts function as temporary scaffolds for the regeneration of the arterial wall.Smooth muscle cells, covering the grafts, regenerated a neo-media underneath an almost completely regenerated endothelial layer (neo-intima). These smooth muscle cells varied in morphology from normal smooth muscle cells to myofibroblasts. They were surrounded by elastic laminae and collagen fibers.Macrophages, epithelioid cells, multinucleated giant cells, fibroblasts and capillaries were present in the disintegrating graft lattices. The epithelioid cells and multinucleated giant cells engulfed polymer particles of the disintegrating grafts.The regeneration of the endothelial and smooth muscle cells is similar to the natural response of arterial tissue upon injury. The presence of macrophages, epithelioid cells, multinucleated giant cells, fibroblasts and capillaries in the graft lattices resembles the natural response of tissue against foreign body implants. Both of these responses result in the formation of a neo-artery that possesses sufficient strength, compliance and thromboresistance to function as a small caliber arterial substitute.Supported by Grant nr. 82.042 from the Dutch Heart Foundation     

Cloning of the pectate lyase genes from Erwinia carotovora and their expression in Escherichia coli

A hybrid cosmid coding for pectate lyase (PL) activity was identified from an Erwinia carotovora genomic library by an immunological screening method. A 7-kb DNA fragment was identified which codes for three proteins identical in size to proteins with PL activity purified from E. carotovora culture supernatants. The three proteins had apparent Mrs of 41, 44 and 44 X 10(3) as estimated by SDS-PAGE. None of the PLs were exported from Escherichia coli strain HB101 but all were found in the periplasmic space. Plant tissue was macerated by the PLs made in E. coli.     

The araBAD operon of Salmonella typhimurium LT2. I. Nucleotide sequence of araB and primary structure of its product, ribulokinase

Hybrid plasmids containing the araBAD operon of Salmonella typhimurium LT2 were characterized by Southern blot and genetic analyses. The nucleotide sequence of araB was determined. The araB gene product, ribulokinase (EC 2.7.1.16), was purified and the results of amino acid composition analysis and partial amino acid sequence are in agreement with predictions from the DNA sequence. Ribulokinase is 569 amino acid residues long and has a calculated Mr of 61 793. Ribulokinase shares significant homology with xylulose kinase from Escherichia coli. Codon usage in the araB gene does not favor those codons which have intermediate codon-anticodon binding energy.     

非洲钝缘蜱感染Q热立克次体敏感度的测定

Q热立克次体以10^-1—10^-12不同稀释度,用血腔注射法感染非洲钝缘蜱,感染35天,取蜱血淋巴液,分别用Gimenez染色法及免疫荧光技术检查,在10^-1—10^-5的稀释度,两种方法检出率基本相似。10^-6以下各组,免疫荧光尚可检出少数阳性标本,而Gimcnez法则无阳性,前者总阳性率为48.28%,后者为40.68%。感染的蜱悬液接种豚鼠,12组动物血清做Q热补体结合试验全部阳性。动物发病情况如潜伏期长短、发烧天数等似与蜱内立克次体的含量有关。     

用电子自旋标记法研究γ辐照对红细胞膜的影响

本文用5-氮氧自由基硬脂酸、12-氮??氧自由基硬脂酸和16-氮氧自由基硬脂酸三种脂肪酸自旋标记物对人红细胞膜(血影)的不同深度膜脂的流动性作了进一步的研究,所得到的电子自旋共振波谱表明它们的序参数都随γ照射剂量的增大而增大。本文还用马来酰亚胺氮氧自由基标记在膜蛋白的SH基团上,所得到的波谱表明其旋转相关时间和强固定化对弱固定化组分的谱线高度的比值也都随γ辐照剂量增大而增大。结果说明γ辐照后,人红细胞膜的流动性降低。这与我们用荧光探针研究所得到的结果相似。