远交系小鼠胚胎干细胞系的建立及嵌合鼠的获得

ＥＳ细胞（ＥｍｂｒｙｏｎｉｃＳｔｅｍＣｅｌｌｓ）是来源于小鼠早期胚胎的多潜能干细胞,它可以在体外大量培养。并以单细胞的形式注射到早期胚胎里,发育为嵌合体。到目前为止,通常使用的１２９小鼠品系是来源于近交系（ｉｎｂｒｅｄ）小鼠的胚胎．与之相比,远交系小鼠应当具有较强的生命力和抗病能力。曾有人报道过建成了远交系小鼠胚胎干细胞系,但是尚没有见到获得嵌合鼠的报道。有人甚至认为：由于不同品系小鼠所具有的遗传背景不同,有的小鼠不能建成ＥＳ细胞系。最近,本实验室在这方面做了有益的探索,成功地建成了远交系小鼠胚胎干细胞系,并在这里报导首例用远交系小鼠胚胎干细胞系培育成功嵌合体小鼠。采用源于Ｓｗｉｓｓ小鼠远交群的昆明（ＫＭ）品系小鼠囊胚建成了三个小鼠胚胎干细胞系（ＫＥ１．ＫＥ２．ＫＥ５）。核型正常率均达到７０％以上。自第八代起分批冻存,复苏后,培养至第１２代,消化成单细胞,通过囊胚显微注射,将其注射到６１５品系小鼠胚胎。在幸存的幼鼠中获得了一只来源于ＫＥ１细胞的嵌合鼠（Ｔａｂｌｅ１）．其毛色表现为受体鼠（６１５）的白色中嵌合有供体鼠（ＫＭ）的黑褐色（ＰｌａｔｅＩ－Ａ）．嵌合鼠与受体鼠的杂交后代鼠中仍然出现了受体鼠的毛色类型（     

地衣芽孢杆菌β－甘露聚糖酶的纯化及酶学性质

地衣芽孢杆菌（Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ）ＮＫ－２７菌株发酵产生的β－甘露聚糖酶（β－ｍａｎｎａｎａｓｅ）经硫酸铵盐析沉淀,两次ＤＥＡＥ纤维素和ＳｅｐｈａｄｅｘＧ－１００离子交换柱层析以及制备ＰＡＧＥ筹步骤,获得了凝胶电泳均一的样品。用ＳＤＳ－凝胶电泳测得纯化后的β－甘露聚糖酶分子量为２６ｋＤ,用凝胶聚焦电泳测得等电点ＰＩ为５．０。酶反应的最适ｐＨ为９．０,最后温度为６０℃,稳定ｐＨ为６．０—９．０,稳定温度为４０℃。金属离子中Ｍｇ￣（２＋）、Ｃａ￣（２＋）、Ｆｅ￣（２＋）、Ｎｉ￣（２＋）对该酶有一定的激活作用;而Ｓｎ￣（２＋）、Ｚｎ￣（２＋）、Ａｌ￣（３＋）、Ａｇ￣＋和Ｈｇ￣（２＋）对该酶有强烈的抑制作用。ＮＫ－２７菌株的β－甘露聚糖酶对魔芋葡萄甘露聚糖和角豆胶半乳甘露聚糖的Ｋｍ值分别为７．１４和５．５６ｍｇ·ｍｌ￣（－１）;Ｖ＿（ｍａｘ）分别为２００．５３和１５７．４５μｍｏｌ·ｍｇ￣（－１）·ｍｉｎ￣（－１）。     

滇产圆滑番荔枝中的番荔枝内酯

从滇南产的圆滑番荔枝的种子中得到４个番荔枝内酯,４－ｄｅｘｏｘｙｃｈｅｒｉｍｏｌｉｎ－２（１）,ｃｈｅｒｉｍｏｌｉｎ－２（２）,ａｎｎｏｎｉｎＩ（３）和ｄｅｓａｃｅｔｙｌｕｖａｒｉｃｉｎ（４）。其中１为新化合物。     

一个Hunter综合征家系粘多糖电泳分析

CT所见肝脏肿大占据左右上腹,纠正了既往把肝左叶当做脾大的结论;B超发现前所没有报道的二尖瓣赘生物将给患者终生致残;标准品DS行双向电泳,首次发现其分离为DS1与GS2两个斑点;杂合子、患者尿GAG均出现DS1斑点,而正常人则不出现。实验显示,DS1的出现在杂合子检出和患者确诊上有重要意义。     

小麦根区的化感物质及其生物活性的研究

小麦根区的化感物质及其生物活性的研究ＳＴＵＤＹＯＮＴＨＥＡＬＬＥＬＯＣＨＭＩＣＡＬＳＰＲＯＤＵＣＥＤＢＹＢＡＣＴＥＲＩＡＡＮＤＦＵＮＧＩＩＮＲＨＩＺＯＳＰＨＥＲＥＡＮＤＴＨＥＩＲＢＩＯＡＣＴＩＶＩＴＹ马瑞霞刘秀芬袁光林孙思恩ＭａＲｕｘｉａＬｉｕＸｉｕ...     

长期施用稀土对小麦植株中稀土元素含量及分布的影响

在我国施用稀土时间最长的黑龙江花园农场,研究了连续１２ａ叶面喷施稀土对小麦植株及土壤中稀土元素总含量和分布模式的影响。结果表明：连续１２ａ叶施稀土没有造成土壤中元素含量及分布模式的变化;对小麦拔节始期（喷施稀土７ｄ）后叶部的影响较大,Ｌａ,Ｃｅ最明显增高,叶部稀土元素分布模式与“常乐”稀土中的相一致,与土壤中稀土元素分布模式不同,而在小麦拔节始期的根部、小麦成熟期的根、茎、叶、壳等部位稀土元素分布     

两种女贞光合作用季节特性研究

以徐州自然条件下生长的常绿女贞和落叶女贞为材料,在不同季节测定了它们的叶绿素含量、光合速率、呼吸速率、光合作用的最适温度。结果表明,两种女贞各测定的生理指标都有季节特性。在自然条件下,两种女贞的光合速率皆为夏季最高,冬季最低,与叶绿素含量变化相一致。在冬季常绿女贞的净光合速率接近于零,而落叶女贞的净光合速率为负值,但仍保持较高的呼吸强度。光合作用的最适温度测定结果表明,常绿女贞适应环境温度变化的能     

Nafion膜固定的亚甲基蓝为介体的生物传感器

制成了以亚甲基蓝为介体的电流型过氧化氢生物传感器,通过离子交换牢固地固定在Nafion膜中的亚甲基蓝,能有效地在辣根过氧化物酶和玻碳电极之间传递电子.探讨了pH值、温度、工作电位和抗坏血酸等物质对此传感器生物电催化还原H₂O₂的影响.此生物传感器选择性好、灵敏度高,对H₂O₂线性响应范围为5.0×10^-7～2×10^-4 mol/L,响应时间少于30 s.     

FK-506结合蛋白对钙释放通道的调控

细胞内自由钙作为一种重要的细胞信使广泛地参与细胞生理功能调控.胞内钙库(内质网系和肌浆网系)对调节细胞内自由钙水平起着重要的作用.钙库膜上的钙释放通道(ryanodine受体和三磷酸肌醇受体)受许多因素调控,其中之一就是新近研究得相当多的FK506结合蛋白.免疫抑制剂FK506能特异地结合钙库上一种分子质量为12 ku左右的蛋白,这种FK506结合蛋白与钙释放通道形成一种紧密连接的复合体,在正常生理情况下对钙释放通道起着十分重要的调控作用.     

Adeno-associated virus type 2-mediated transfer of ecotropic retrovirus receptor cDNA allows ecotropic retroviral transduction of established and primary human cells.

The cellular receptors that mediate binding and internalization of retroviruses have recently been identified. The concentration and accessibility of these receptors are critical determinants in accomplishing successful gene transfer with retrovirus-based vectors. Murine retroviruses containing ecotropic glycoproteins do not infect human cells since human cells do not express the receptor that binds the ecotropic glycoproteins. To enable human cells to become permissive for ecotropic retrovirus-mediated gene transfer, we have developed a recombinant adeno-associated virus type 2 (AAV) vector containing ecotropic retroviral receptor (ecoR) cDNA under the control of the Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter (vRSVp-ecoR). Established human cell lines, such as HeLa and KB, known to be nonpermissive for murine ecotropic retroviruses, became permissive for infection by a retroviral vector containing a bacterial gene for resistance to neomycin (RV-Neo(r)), with a transduction efficiency of up to 47%, following transduction with vRSVp-ecoR, as determined by the development of colonies that were resistant to the drug G418, a neomycin analog. No G418-resistant colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. Southern and Northern blot analyses revealed stable integration and long-term expression, respectively, of the transduced murine ecoR gene in clonal isolates of HeLa and KB cells. Similarly, ecotropic retrovirus-mediated Neo(r) transduction of primary human CD34+ hematopoietic progenitor cells from normal bone marrow was also documented, but only following infection with vRSVp-ecoR. The retroviral transduction efficiency was approximately 7% without prestimulation and approximately 14% with prestimulation of CD34+ cells with cytokines, as determined by hematopoietic clonogenic assays. No G418-resistant progenitor cell colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. These results suggest that sequential transduction of primary human cells with two different viral vectors may overcome limitations encountered with a single vector. Thus, the combined use of AAV- and retrovirus-based vectors may have important clinical implications for ex vivo and in vivo human gene therapy.