中国人肥胖基因真核表达载体的构建

为研究肥胖（ｏｂｅｓｉｔｙ）的病因及肥胖基因（ｏｂ）的表达与调控,根据文献报道的ｈｏｂ序列设计引物,经ＲＴ－ＰＣＲ扩增中国人的ｏｂ基因（包括信号肽在内的ｃＤＮＡ全长５４０ｂｐ）,ＰＣＲ产物采用Ｔ－Ａ克隆法首先连接到克隆载体ｐＵＣ１１９,然后定向转移到经改造的真核表达载体ｐＳＶ－β－ｌａｃＺ,酶谱分析表达克隆基因为的人肥胖基因。     

生物信息学技术克隆人类神经髓鞘蛋白零家族基因

为克隆新的髓鞘蛋白相关基因,将ＭＰＺ基因编码区ｃＤＮＡ与ＥＳＴ数据库进行同源性比较,得到与ＭＰＺ显著相似的２个ＥＳＴ,构建成８０１ｂｐ的重叠群。此重叠群与定位在１ｑ２４的１２８ｋｂ ｇＤｎＡ的相似性为１００％。计算机分析有ｂｐ和重叠群可能存在一个４３５ｂｐ的阅读框架。在重叠群上设计两个引物与文库载体臂上的引物配对,扩增各种ｃＤＮＡ文库ＤＮＡ作巢式ＰＣ拓所得ｃＤＮＡ上再设计引物进行巢式ＰＣＲ,最终克     

重叠片段法克隆全长人肝细胞生长因子cDNA

通过重叠片段的ＲＴ－ＰＣＲ方法,从人胎盘组织克隆了人全长肝细胞生长因子ｃＤＮＡ片段（约２２００ｂｐ）,经酶切证实和测序分析都表明该片段确为人ＨＧＦｃＤＮＡ。本文所用方法解决了扩增较长目的基因片段时,由于ＲＮＡ酶及反转录酶的ＲＮＡ酶Ｈ活性和ｍＲＮＡ二级结构等多种因素的影响,使获取长片段ｃＤＮＡ难以成功的难点     

从9q34肌氨酸血症位点克隆人二甲基甘氨酸脱氢酶样基因

将鼠二甲基甘氨脱氨酶样基因（ｉｍｅｔｈｙｌｇｌｙｃｉｎｅｄｅｈｙｄｒｏｇｅｎａｓｅ－ｌｉｋｅｇｅｎｅ）的部分 编码序列对ＥＳＴ数据库进行同源性分析,得到与其显著相似的１个人ＥＳＴ（ＧｅｎＢａｎｋＨ２８８５６）（３３０ｂｐ）。此ＥＳＴ与９ｑ３４上的２个ｇＤＮＡ的文库中载体臂上上两端的引物ＰＣＲ,在所得ｃＤＮＡ上再设计引物与ｃＤＮＡ文库载体臂上引物ＰＣＲ,最终在肝ｃＤＮＡ文库获得１个完整编码区的ｃＤＮ     

芸苔属植物CAL基因的结构特征与花球的形态遗传

从花椰菜变种（Ｂｒａｓｓｉｃａ ｏｌｅｒａｃｅａ Ｌ．ｖａｒ．ｂｏｔｒｙｔｉｓ）中分离出４种花球类型的纯合株系：光滑型花球ｃｕｒｄ－ｓ、毛状物花球ｃｕｒｄ－ｈ、颗粒状花球ｃｕｒｄ－ｃ和刺状物花球ｃｕｒｄ－ｔ。用ＰＣＲ方法分别扩增各株系的ＢｏｂＣＡＬ基因并分析其中的部分序列,发现４种株系的ＣＡＬ基因在第五个外显子中都有ＡＡＧ向ＴＡＧ的终止突变,而且该终止密码子上游６３８ｂｐ的ＤＮＡ序列完全一致,说明     

热休克后表达受抑基因的分子克隆及其表达特性

以人成骨肉瘤细胞株ＨＯＳ－８６０３为热休克模型,在利用ＤＤｍＲＮＡ方法筛选和克隆了一个热休克后表达受抑基因的ｃＤＮＡ片段的基础上,以该片段为探针,筛选ＨＯＳ－８６０３细胞的ｃＤＮＡ文库,获得该基因的全长ｃＤＮＡ克隆（ＨＳＳＧ－１）;ＤＮＡ序列测定表明该ｃＤＮＡ全长１４５６ｂｐ,编码２７６个氨基酸,经计算机辅助分析,该ｃＤＮＡ序列尚未被报道。经ＲＮＡ斑点杂交证实,该基因的表达于多种组织细胞之中,并且     

大鼠脑谷氨酸脱羧酶基因的cDNA克隆及序列分析

胰岛β－细胞自身抗原蛋白之一是脑中谷氨酸脱羧酶（Ｇｌｕｔａｍｉｃａｃｉｄｄｅｃａｒｂｏｘｙｌａｓｅ,ＧＡＤ,ＥＣ４．１．１．１５）同源物。以双链ｃＤＮＡ为模板,用ＰＣＲ方法快速克隆了Ｗｉｓｔａｒ大鼠脑ＧＡＤ基因的ｃＤＮＡ,将此包括编码５９３个氨基酸的全长ＤＮＡ片段重组入ｐＵＣ质粒并用双脱氧末端终止法测定了全部序列,证明其全长为１７７９ｂｐ．经比较发现Ｗｉｓｔａｒ大鼠脑与Ｒｕｓｓｅｌｌ报导的大鼠脑ＧＡＤ基因序列,有一处碱基的差别,但并不涉及氨基酸的改变。同时还对用ＰＣＲ扩增长片段ＤＮＡ进行了方法学上的探讨。     

一个新的水稻MADS—box基因的克隆及表达分析

根据ＭＡＤＳ－ｂｏｘ基因保守区结构,设计简并性引物,利用３＇ＲＡＣＥ从水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）中克隆了１个新的水稻ＭＡＤＳ－ｂｏｘ基因的ｃＤＮＡ片段,同时利用５＆ＲＡＣＥ获得了全长ｃＤｎＡ命名为ＦＤＲＭＡＤＳＳ。序列分析表明,该ｃＤＮＡ全长１４０６ｂｐ,开放阅读框共编码２３３个到,具有典型的植物ＭＡＤＳ－ｂｏｘ基因的结构。推测的氨基酸序列与拟南芥的ＭＡＤＳ－ｂｏｘ基因,ＡＧＬ１４同     

PCR——四步两段扩增法分离人碱性成纤维细胞生长因子（hbFGF）基因

引物是决定ＰＣＲ结果的关键,本实验中,引物Ⅰ,Ⅱ虽有２６个碱基,但由于其５′端有８个碱基为限制性内切酶的识别序列,所以实际上只有１８个碱基与原始模板互补配对,采用标准的ＰＣＲ方法时不能产生理想结果,经采用ＰＣＲ－四步两段扩增法从人胎盘ｃＤＮＡ文库中分离得到了－４８４ｂｐ的ＤＮＡ片段。通过斑点杂交鉴定,结果表明该ＤＮＡ片段为ｈｂＦＧＦ基因。     

大鼠脑谷氨酸脱羧酶基因的cDNA克隆及序列分析

胰岛β－细胞自身抗原蛋白之一是脑中谷氨酸脱羧酶（Ｇｌｕｔａｍｉｃａｃｉｄｄｅｃａｒｂｏｘｙｌａｓｅ,ＧＡＤ,ＥＣ４．１．１．１５）同源物,以双链ｃＤＮＡ为模权,用ＰＣＲ方法快速克隆了Ｗｉｓｔａｒ大鼠脑ＧＡＤ基因的ｃＤＮＡ将此包括编码５９３个氨基酸的全长ＤＮＡ片段重组入ｐＵＣ质粒并用双脱的氧末端终止法测定了全部序列,证明其全长为１７７９ｂｐ,经比较发现Ｗｉｓｔａｒ大鼠脑与Ｒｕｓｓｅｌｌ报导的大鼠脑Ｇ     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

Identification and Characterization of the First Equine Parainfluenza Virus 5

正Dear Editor,Parainfluenza virus 5 (PIV5), known as canine parainfluenza virus in the veterinary field, is a negative-sense,nonsegmented, single-stranded RNA virus belonging to the Paramyxoviridae family (Chen 2018). The virus was first reported in primary monkey kidney cells in 1954 (Hsiung1972), then it has been frequently discovered in various     

Pathogenic Characterization and Full Length Genome Sequence of a Reassortant Infectious Bursal Disease Virus Newly Isolated in Pakistan

<正>Dear Editor,Infectious bursal disease (IBD) is one of the most important diseases of the poultry. The IBD virus (IBDV), a nonenveloped virus belonging to the Birnaviridae family with a genome consisting of two segments of double-stranded RNA (segments A and B), targets B lymphocytes of bursa of Fabricious leading to immunosuppression. In Pakistan,poultry farming is the second biggest industry and IBD is the second biggest disease threating the poultry sector.However, there is limited genome information of IBDV     

Similar aerosol emission rates and viral loads in upper respiratory tracts for COVID-19 patients with Delta and Omicron variant infection

Highlights
1 Aerosol emission rates of Delta or Omicron patients were similar.
2 Viral loads in upper respiratory tract of Alpha, Delta and Omicron patients were similar.
3 Viral loads in upper respiratory tract of vaccinated or unvaccinated Delta patients had no difference.     

Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b in vitro

Highlights
1) A comprehensive evaluation method for anti-SARS-CoV-2 drugs was established based on RT-qPCR, TCID₅₀ method, and immunofluorescence.
2) A significant antiviral effect of rHuIFN-α1b was shown with EC₅₀=0.12 IU/mL in Vero cells and EC₅₀=0.52 IU/mL in Calu-3 cells, which was better than rHuIFN-α2b (EC₅₀=0.25 IU/mL in Vero cells and EC₅₀=2.48 IU/mL in Calu-3 cells).
3) rHuIFN-α1b has a good potential in the application of anti-COVID-19 therapy.