A SPOROPHYTE CELL WALL PROTEIN OF THE RED ALGA PORPHYRA PURPUREA (RHODOPHYTA) IS A NOVEL MEMBER OF THE CHYMOTRYPSIN FAMILY OF SERINE PROTEASES1

A complementary DNA (cDNA) clone from a Porphyra purpurea (Roth) C. Agardh sporophyte-specific subtracted cDNA library was found to encode a protein similar to serine proteases of the chymotrypsin class. The encoded protein contains a typical signal peptide and is particularly similar to chymotrypsins in the regions surrounding the active site residues and the activation site where cleavage of the propeptide occurs. In addition, the six cysteine residues characteristic of chymotrypsins are conserved. However, two of the three residues of the active site His/Asp/Ser charge relay triad have been replaced, indicating that the protein is unlikely to have peptidase activity. Northern hybridization confirmed that this cDNA is derived from an abundant, sporophyte-specific messenger RNA (mRNA). The presence of signal peptide on the encoded protein and the abundance of its mRNA suggested that this protein might be localized in the cell wall. Consequently, sporophyte cell walls were isolated and a major protein having a molecular weight similar to that estimated for the encoded protein was purified. N-terminal sequence analysis indicated that this cell wall protein is identical to that encoded by the cDNA with the amino terminus of the mature protein beginning at the activation site. This cell wall structural protein appears to have evolved from a chymotrypsin-like progenitor but has been adapted to bind cell wall proteins and/or polysaccharides rather than to cleave proteins.     

细胞分裂素生物合成基因在调节一组烟草病理相关蛋白基因表达中的作用

对一组病理相关蛋白基因在烟草 ( N icotiana tabacum cv. Wisconsin 38)中的表达情况进行了研究 ,包括 :碱性几丁质酶、β- 1 ,3-葡萄糖苷酶、渗透蛋白及伸展蛋白。RNA杂交实验表明在正常烟草植株中上述 4个基因具有发育和器官专一性的表达。在含有细胞分裂素生物合成基因的转基因烟草丛生芽中 ,这 4个基因的表达受过量合成的内源细胞分裂素和载体效应的共同调节 ,细胞分裂素降低这些基因的表达 ,而载体效应则促进它们的表达。热激处理也明显降低这 4种基因的表达水平。上述结果表明这些病理相关蛋白基因具有复杂的调控系统     

Elongation factor 1α genes of the red alga Porphyra purpurea include a novel, developmentally specialized variant

The life cycle of the red alga Porphyra purpurea alternates between two morphologically distinct phases: a shell-boring, filamentous sporophyte and a free-living, foliose gametophyte. From a subtracted cDNA library enriched for sporophyte-specific sequences, we isolated a cDNA encoding an unusual elongation factor 1 (EF-1) that is expressed only in the sporophyte. A second EF-1 gene that is expressed equally in the sporophyte and the gametophyte was isolated from a genomic library. These are the only EF-1 genes detectable in P. purpurea. The constitutively expressed gene encodes and EF-1 very similar to those of most eukaryotes. However, the sporophyte-specific EF-1 is one of the most divergent yet described, with nine insertions or deletions ranging in size from 1 to 26 amino acids. This is the first report of a developmental stage-specific EF-1 outside of the animal kingdom and suggests a fundamental role for EF-1 in the developmental process.     

应用MUCAP试剂快速检测沙门氏菌

报告了用4-甲基伞形酮辛酯(4-Methylumbelliferyl-caprylate, MUCAP)快速检测沙门氏菌的特异性、敏感性和实用性。经HE,DHL,SS和麦康凯琼脂平板分离的65株沙门氏菌标准菌株和48株从食品中分离的沙门氏菌,用MUCAP测试均呈阳性反应;394株非沙门氏菌中呈阳性反应的假单胞菌、气单胞菌、邻单胞菌可通过氧化酶试验与沙门氏菌区分开;与粘质沙雷氏菌的交叉反应改用加1％蔗糖的分离平板也可排除。此方法的敏感性和特异性均达到97％以上,而且操作简便、快速,数分钟内即可完成。     

有机硅橡胶固定化细胞进行的生物转化

有机硅橡胶固定化细胞进行的生物转化潘冰峰,戴学倩,冯青,李祖义（中国科学院上海有机化学研究所,２０００３２）关键词硅橡胶;白地霉;固定化细胞;生物转化近年来,有机化学领域的一个重要进展是用酶或微生物进行生物转化反应。其中研究和应用较多的是碳基还原,特...     

利福霉素生产菌产生钝化RifSV物质的分离及性质研究

利福霉素ＳＶ（简称ＲｉｆＳＶ）生产菌──地中海拟无枝菌酸菌在生物合成ＲｉｆＳＶ过程中,产生一种能钝化自身产物（ＲｉｆＳＶ）的物质．实验证实,该物质是由谷氨酸、天冬氨酸、赖氨酸及缬氨酸等１４种常见氨基酸组成的蛋白质．分子量（ＭＷ）约２．５×１０４Ｄ,等电点（ＰＩ）为５．７—６．１．在１００℃下加热１０ｍｉｎ,其活性丧失．作用于ＲｉｆＳＶ的最适ｐＨ值范围为７．４—８．６,最适温度为２９℃,初步证实该物质是一种酶（暂称利福霉素钝化酶）     

人白介素6受体功能区片段在E．coli中的表达

通过ＤＮＡ体外重组技术,以ｐＥＴ－３ｂ为表达载体,构建了重组表达质粒ｐＥＴ－６Ｒ（Ｂ）和ＰＥＴ－６Ｒ（Ｂ）４,分别编码２８ｋＤ的ｈＩＬ－６Ｒ配基结合区片段及其５３ｋＤ的二联体蛋白,并为酶切分析和ＤＮＡ序列分析所证实。ＳＤＳ－ＰＡＧＥ分析表明,含有重组表达质粒的菌株可分别表达出２８ｋＤ的蛋白ｒＩＬ６Ｒ－２８和５３ｋＤ的ｒＩＬ６Ｒ－５３。重组蛋白分别占菌体总蛋白的４５％和２９％左右。重组蛋白主要以包涵体形式存在,Ｗｅｓｔｅｒｎ印迹表明重组蛋白具有ＩＬ－６Ｒ的抗原性。     

汉防己甲素及克矽平对矽肺大鼠Ⅰ型和Ⅲ型胶原mRNA的影响

汉防己甲素（汉甲）及克矽平（Ｐｏｌｙｖｉｎｙｌｐｙｒｉｄｉｎｅ－Ｎ－Ｏｘｉｄｅ,ＰＶＮＯ）是目前较为有效的抑制矽肺纤维化的药物。本文研究了其对胶原ｍＲＮＡ水平的影响．斑点杂交实验表明大鼠接尘６０天和１２０天后α１（Ⅰ）及α１（Ⅲ）ｍＲＮＡ水平明显上升,经汉甲或克矽平治疗１个月或３个月后,胶原ｍＲＮＡ水平明显下降。原位杂交结果表明胶原ｍＲ－ＮＡ银颗粒与细胞性结节和增厚的肺泡壁的成纤维细胞分布重合。汉甲或克矽平治疗后银颗粒数下降。提示汉甲及克矽平对矽肺进程中的胶原基因表达增强有抑制作用。     

天麻中抗真菌蛋白质的诱导和积累

前曾报告从天麻（Ｇａｔｒｏｄｉａ ｅｌａｔａ Ｂｌ）中分离出一种对木霉丝生长有强抑制作用的蛋白质,命名为天麻抗真菌蛋白,简称ＧＡＦＰ,也称为Ｇａｓｔｒｏｄｉａｎｉｎ。对不同天麻材料来源的ＧＡＦＰ分析表明：ＧＡＦＰ的相对分量为１４ｋＤ,等电点可能不同,变动范围８．１－９．３。体外抑菌试验证明ＧＡＦＰ对腐生性真菌如木霉如木霉和密环菌等有强抑制活性,半抑制浓度ＩＣ０．５为０．０８ｍｇ／ｍＬ;对寄生免疫荧     

新疆地区小麦白粉病菌寄主范围的研究

用小麦白粉病菌11个生理小种的混合菌种,对新疆地区的小麦近缘植物的7个属22个种的47份材料进行接种,除6份免疫外,其余均接种成功.用其中6个属19个种的29份小麦近缘植物产生的白粉病菌,对小麦回接,参试的29份材料全部回接成功.小麦白粉病菌对小麦近缘植物的寄生像在小麦上一样,有明显的寄生专化性.感病的小麦近缘植物的78.0%对小麦白粉病菌的感病性,随生育期增长而急剧下降.文中并对小麦白粉病中间寄主的作用进行了讨论.