中国人HLA-G1的cDNA克隆、序列分析、表达和蛋白质空间结构预测

从中国人外周血单个核细胞、胎盘组织和肝癌组织等样品中克隆了包含完整HLA-G1读框的cDNA;与国外同行获得的该基因及其蛋白质序列比较分析表明,该基因虽然有着细微的种族特异性,但高度保守;并获得了它的截断型重组蛋白,根据蛋白一级结构和同源比较方法,模建了它及其与特异性受体KIR2DL4形成复合体的空间结构模拟,预测了它们之间相互作用的特征.     

Construction of human and mouse brain cDNA libraries and isolation of full-length cDNAs

cDNA libraries from aborted human 3-month fetal brain,adult rat and mouse brain were constructed by using a yZAP express cDNA library construction kin.Low molecular weight fragments of the second strand cDNASA were removed by flowing through the Sepharose CL-4B column and the frractionated long,Middle,Short fragments and the combined fragments weire respectively inserted into clone vectors to construct the cDNA libraries of the brain of human 3-month fetus.The 5'ends of 1200 clones from each of human fetal brain cDNA libraries were sequenced.A total of 894 ESTs were obtained and some full-length clones were squenced.By andalyaing the se-quences,12 novel full-length cDNAs were obtained.     

旋毛虫新生幼虫cDNA文库的构建与鉴定

利用λZAPExpress载体成功地构建了中国猪株旋毛虫分离株Trichinellaspiralis新生幼虫的cDNA文库 ,并对其重组噬菌体质粒pBK—CMV进行酶切鉴定 .结果表明 :所构建的cDNA文库容量为 1 92× 10 6,重组效率为 98 6 % ,所有的克隆片段都在 0 5— 2 0Kb之间 ,说明此cDNA文库几乎覆盖了全部mRNA .     

人类STK全长cDNA克隆及其表达谱分析

从人类的睾丸组织ｃＤＮＡ文库中分离出１条新的全长ｃＤＮＡ,在其１２２５ｂｐ序列中含有一个长１０３２ｂｐ,编码３４４个氨基酸的开放阅读框。基于它与丝／苏氨酸蛋白激酶家族中其他成员存在较高的同源度,尤其是与小鼠ＭＭＳＴＫ１的同源度高达８０％,推论该基因的蛋白产物可能也是一个丝／苏氨酸蛋白激酶,故将其暂命名为ＨＵＭＳＴＫ１（人类ＳＴＫ１）。它的ｍＲＮＡ在胎儿胸腺中呈高表达,在睾丸、小肠和结肠中低表达,而     

Infectious foot-and-mouth disease virus derived from a cloned full-length cDNA of OH／CHA／99

The China foot-and-mouth virus (FMDV) isolate OH/CHA/99 was isolated from swine, which was unable to infect bovine thyroid cells in vitro or to cause typical disease in bovines following intradermal inoculation in the tongue. To enhance antigenicity, replication, maturation and pathogenicity studies of OH/CHA/99, an infectious fulllength cDNA clone, designated pBIFMDV, was prepared. The in vitro and in vivo biological properties of the virus derived from pBIFMDV were studied by analyzing antigenicity, plaque morphology and virulence in pigs. The results showed that the virus derived from pBIFMDV had the same biological properties as the parent strain OH/CHA/99; the fulllength infections cDNA clone, pBIFMDV, will be very useful in studies of the antigenicity, virulence, pathogenesis, maturation and replication of FMDV.     

Notable diversity in hemoglobin expression patterns among species of the deep-sea clam, <Emphasis Type="Italic">Calyptogena</Emphasis>

The deep-sea clams Calyptogena nautilei and C. tsubasa, which live in the cold-seep area at a depth of 3570 m in the Nankai Trough, Japan, have abundant hemoglobins (Hbs) in erythrocytes, similar to other Calyptogena species. We determined the cDNA-derived amino acid sequences of Hbs from two Calyptogena species. C. tsubasa was found to contain two dimeric Hbs, Hb I consisting of 145 amino acid residues and Hb II with 137 residues, similar to known Hbs from C. soyoae and C. kaikoi. Sequence identity was over 90% among the orthologous chains of Calyptogena Hbs. On the other hand, surprisingly, C. nautilei contained two monomeric Hbs, Hb III containing 141 residues and Hb IV with 134 residues. In addition, Hbs III and IV showed only 33–42% sequence identity with Hbs I and II from other Calyptogena species. The distal (E7) histidine, one of the functionally important residues of the heme protein, is replaced by glutamine in all Hb chains of Calyptogena species. A phylogenetic analysis indicated that C. nautilei Hb III is closer to Hb I from other Calyptogena species. We suppose that a Hb gene was duplicated at least three times in an immediate ancestor of Calyptogena and, presumably depending on physiological conditions different Hb sets are being expressed: dimeric Hbs I and II in C. soyoae, C. kaikoi and C. tsubasa, and monomeric Hbs III and IV in C. nautilei. Received 13 May 2003; received after revision 5 June 2003; accepted 12 June 2003     

中华绒螯蟹卵巢新基因EJO6的全长cDNA克隆和序列分析

利用RACE技术从中华绒螫蟹卵巢获得了新基因EJO6(Eriocheir japonica ovary gene 6,EJO6)的全长cDNA序列(GenBarnk 检索号：AYl85922)。该cDNA序列长度为1250bp,开放阅读框为690bp,编码229个氨基酸。根据氨基酸序列计算的相对分子质量和等电点分别为24270和11．9。同时用生物信息学的方法对该基因的结构和功能进行了初步分析。     

抗白粉病的小麦-黑麦1BL/1RS易位系99/2439的cDNA文库构建

用pGEMT-easy质粒载体构建了经白粉茵诱导0.5h、1.0h、12h、24h和48h的小麦-黑麦1BL/1RS易位系99/2439叶片cDNA文库1个.文库宿主茵为E. coliDH10B,诱导文库平均插入片段为1kb左右.     

Magnaporthe oryzae MTP1 gene encodes a type III transmembrane protein involved in conidiation and conidial germination

In this study the MTP1 gene, encoding a type III integral transmembrane protein, was isolated from the rice blast fungus Magnaporthe oryzae. The Mtp 1 protein is 520 amino acids long and is comparable to the Ytp 1 protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP （green fluorescent protein） fusion expression results indicate that Mtp 1 is a protein located at several membranes in the cytoplasm. The functions of the MTP1 gene in the growth and development of the fungus were studied using an MTP1 gene knockout mutant. The MTP1 gene was primarily expressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for pathogenicity. The Amtpl mutant grew more efficiently than the wild type strain on non-fermentable carbon sources, implying that the MTP1 gene has a unique role in respiratory growth and carbon source use.