Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

汕头市松材线虫病的防治

基于汕头市濠江区已发现松材线虫病疫情的实际情况,针对该病的防治特点,结合汕头市各辖区山林的实际情况,从判断、预防、治理三个方面提出相应的防治对策,为科学防控该病在我市的扩散危害提供参考。     

奶牛乳房炎多联苗(B)的临床免疫试验

用奶牛乳房炎多联苗 (B)免疫家兔 ,14 d血清中金黄色葡萄球菌抗体效价可达 1∶ 8,2 8d抗体水平明显升高 ,可达 1∶ 32 ,注苗后 35 d攻毒 ,攻毒后 2 d保护率为 93.3% (14 /15 ) ,自然保护率为 2 0 % (1/5 ) ,10 d保护率为 6 6 .6 % (10 /15 ) ,自然保护率为 2 0 % (1/5 ) ;用多联苗 (B)臀部肌肉注射免疫泌乳牛 ,试验期间 4个月内 ,3ml剂量组临床型乳房炎月平均发病率为 11.0 9% ,其对照组为 2 0 .19% ,发病率降低 4 5 .2 9% ,差异极显著 (P<0 .0 1) ;5 m l剂量组月平均发病率为 13.0 1% ,其对照组为 31.36 % ,发病率降低 5 8.5 1% ,差异极显著 (P<0 .0 1) ;3ml剂量组与 5 m l剂量组之间发病率无明显差异 (P>0 .0 5 ) ;注苗后泌乳牛血清抗体水平可持续 4个月 ,且 30 d时效价水平最高。     

催干剂对氨基环氧树脂阴极电泳漆的影响

以氨基环氧树脂为基体树脂，Co^2 ,Pb^2 盐为催干剂合成了阴极电泳漆，并对膜性能进行考察，结果表明，所合成的阴极电泳涂料在PH值为6－7、电泳时间2min的条件下。加入不同的催干剂，在不同的烘干温度下，得到性能不同的漆膜。经过膝膜综合性能试验，结果表明，加入钴和铅复合催化剂，在160℃左右烘干20min，可得到完全固化，硬度较高、耐潮、耐盐、耐酸碱性较好的漆膜。     

猪血浆蛋白(酶)多态性与杂种优势的关系

为探讨血浆蛋白 (酶 )多态性与杂种优势的关系 ,测定了杜洛克、长白、大白、杜×长大、大×长大、长×大、大×长共 7个品种 (组合 )的 8个血浆蛋白 (酶 )位点的多态性及部分生长和胴体性状 ,计算了平均基因杂合度与部分经济性状实测值和杂优率的相关关系 .结果表明 ,平均基因杂合度与遗传距离呈正相关 ,与日增重、屠宰率、背膘厚、后腿比例的实测值或杂优率呈正相关 ,与眼肌面积的实测值和杂优率呈负相关 .平均基因杂合度和亲本间遗传距离可为预测杂种优势提供依据 .     

对《中图法》(第四版)类目注释的系统认识

本文从语言特点、编排体例及内容特点出发,对《中图法》(第四版)类目注释进行了系统的认识,并就此提出了自己的一些看法。     

基于果实相关性状的桃品种初级核心种质取样策略研究

以国家种质资源圃(北京)编目的558份桃品种的18个形态学和农艺学性状为基本数据,研究了桃品种初级核心种质构建的取样策略,包括总体取样比例的确定及取样方案的选择,以获得最佳的初级核心种质。本试验共设10%～90%9个总体取样比例;取样方案研究包括分组原则和组内取样比例的确定。结果表明,桃初级核心种质的适宜总体取样比例为10%;按品种类群分组结合多样性比例取样为构建桃初级核心种质的最佳取样方案;利用此取样策略从558份桃品种中提取56份作为核心样本,对其代表性进行检测表明所构建的初级核心种质能够很好地代表桃原始种质的遗传多样性。     

离子选择电极法测定NBPG与银、镉、汞的配合物组成和稳定常数

本文用离子选择电极法测定了N,N—二甲基膦酸基甘氨酸(NBPG)与银、镉、汞的配合物组成和稳定常数。结果表明,NBPG与Ag~ 、Cd~(2 )、Hg~(2 )均生成组成为1:1的多齿质子型配合物。其lgK_稳值分别为:AgL5.28、CdH_3L3.63、CdH_2L5.16、CdHL7.30、CdL11.56、HgH_3L6.06、HgH_2L7.55、HgHL8.24、HgL12.58。     

应用生物素标记的NDV－cDNA探针检测感染尿囊液中NDV－RNA的初步研究

本文应用聚合酶链反应（ＰＣＲ）技术从构建的新城疫病毒（ＮＤＶ）ｃＤＮＡ文库中扩增含编码Ｆ糖蛋白前体──Ｆｏ酶切位点序列的３５９ｂｐ的Ｆ蛋白基因ｃＤＮＡ片段。将此３５９ｂｐｃＤＮＡ片段经光敏生物素标记后,即成ＮＤＶ－ｃＤＮＡ探针。该探针能特异性地从感染的尿囊液中检测出ＮＤＶ强毒株和疫苗毒株的基因组ＲＮＡ,而不与ＩＢＤｖ－ｄｓＲＮＡ、ＡＩＢｖ－ｓｓＲＮＡ、ＥＤＳ７６－ｄｓＤＮＡ、ＭＤＶ－ｄｓＤＮＡ,ＦＰＶ－ｄｓＲＮＡ及ＡＩＬＶ－ｄｓＤＮＡ发生交叉杂交反应。试验结果表明：尽管该探钎含有编码Ｆｏ蛋白酶切位点序列的碱基顺序,但它还是不能把ＮＤＶ的强、弱毒株区分开。这说明ＮＤＶ强、弱毒株比区域内的碱基存在着相当大的同源性。不过,此探针对ＮＤＶ来说具有特异性,这就为ＮＤＶ的诊断技术开创了基因水平检测的新途径。     

Unsaturated vitamin b(12) binding capacity in human and ruminant blood serum - a comparison of techniques including a new technique by high performance liquid chromatography

A new technique by High Performance Liquid Chromatography (HPLC-gel permeation) shows promise as a tool to separate and quantitate the Unsaturated Vitamin B(12) Binding Capacity (UBSC) of the individual Vitamin B(12) binders in blood serum. This method, although not as rapid as protein-coated charcoal or cellulose separation techniques, is more applicable for use with large numbers of samples than gel filtration. The use of a radioactivity detector to monitor the eluant from the column permitted automation of the method. Comparable results for UBBC and for the UBBC of individual binders were obtained when samples were analyzed by gel filtration and HPLC. The HPLC method proved suitably precise and the recovery of added cyanocobalamin was acceptable. It is proposed that HPLC be the method of choice for measurement of the USBC of binders of Vitamin B(12) in blood serum.