盐酸胍诱导的α淀粉酶去折叠与再折叠

利用蛋白质内源荧光和酶活性两种信号以及荧光偏振,HPLC和停流等方法研究了盐酸胍诱导的α淀粉酶去折叠与重折叠的平衡转变和动力学。实验结果表明α淀粉酶去折叠与重折叠是两个不同的过程;变性与复性过程中可能伴有聚集体生成;去折叠与重折叠均为双相过程,重折叠大约始于2秒之后。     

大鼠胰腺γ-氨基丁酸免疫组织化学定位

本文用ABC—GDN免疫组织化学方法,研究了γ-氨基丁酸(Gamma—Aminobutyric Acid,GABA)在大鼠胰腺的定位和分布,并用相邻切片法,观察它与胰岛素的共存关系。结果发现GABA免疫反应阳性细胞主要分布于胰腺内分泌部(胰岛)。在外分泌部亦有少许分布。大部分胰岛细胞呈GABA免疫反应阳性,集中位于胰岛的中央部。相邻连续切片免疫染色证实GABA与胰岛素共存于胰岛B细胞中。外分泌部胰腺GABA免疫反应阳性细胞,呈零散分布于腺泡和导管上皮间。本文为进一步探讨GABA在胰腺的生理作用提供了形态学依据。     

EFFECTS OF VASOACTIVE INTESTINAL PEPTIDE ON ACTIVITIES OF SUCCINIC DEHYDROGENASE AND ALKALINE PHOSPHATASE IN RAT HEPATOMA CELLS

Using cytochemical method,microspectrophotometry and image analysis,effects of va-soactive intestinal peptide(VIP)on activities of succinic dehydrogenase(SDH)and alkalinephosphatase(ALP)in rat hepatoma cells were studied in vitro.The results showed that thehepatoma cell expressed potent positive reactions of SDH and ALP,the positive positionswere located at the cell membranes and/or cytoplasm.Having been treated with VIP,ALPdecreased obviously in activity(P<0. 01,compared with hepatoma cells untreated by VIP).The sites of ALP activty were chiefly located at the cell membranes,particularly at the cell-cell contacts.Cultured rat hepatoma cells had intensive SDH activity in their cytoplasm.Compared with untreated eclls,there was no marked difference in the intensity of SDH activ-ity in VIP-treated hepatoma cells(P>0.05).     

赤子爱胜蚯蚓(Eisenia Foetida)纤溶酶的研究,Ⅳ纤溶酶组成成分的分子量测定

本文报告了用SephadexG—100柱层析法纯化样品和聚丙烯酰胺凝胶电泳测定纤溶酶组成成分的分子量的研究结果。经柱层析分离的纤溶酶电泳测定分子量结果为11条带,分别为68,000、49,000、42,000、41,000、36,000、29,000、27,000、26,000、25,000、21,000和12,000。而纤溶酶的主要组分集中在21.000与42.000之间,为其活性组分。     

蛋白质结构成对比较的新方法

介绍一种蛋白质三维结构的快速比较方法.此方法毋需初始联配,而能自动寻找和智能迭代.利用本程序对珠蛋白、丝氨酸蛋白酶、天冬氨酸蛋白酶、钙结合蛋白和溶菌酶作了系统的结构比较,取得了满意的结果.本文也讨论了衡量联配结果好坏的要素问题.     

Redescription of the Chengjiang arthropod Squamacula clypeata Hou and Bergström, from the Lower Cambrian, south-west China

The anatomy of the arthropod Squamacula clypeata Hou and Bergström, 1997 from the Lower Cambrian Chengjiang Lagersta¨tte is redescribed based on four newly excavated specimens. The new material was collected from localities recently discovered in the Kunming area, Yunnan Province, south-west China, and preserves remarkable details of the ventral morphology, revealed by preparation. Squamacula clypeata is dorsoventrally flattened and rounded in outline. The cephalon was covered by a wide, short shield, with a large doublure and a pair of uniramous antennae on the ventral side. The thorax consists of nine somites, each protected by a tergite and carrying at least one pair of biramous limbs. The pygidium is covered with a small rounded tergum. The endopod is segmented, equipped with short spines on the inner margin of the coxa and a claw-like structure distally, and the exopod flap-like, fringed with setae. The limbs in the pygidium are like those in the thorax in shape. Squamacula was most probably a nektobenthic predator. The spinose endopod could walk, grasp and grind. The large flap-like exopod was adapted for swimming and respiration. Its affinities lie with the Arachnomorpha, but the relationships with other known taxa remain ambiguous.     

The proteins encoded by the rbs operon of Escherichia coli: II. Use of chimeric protein constructs to isolate and characterize RbsC.

Chimeric genes encoding full-length copies of rbsA and rbsC connected by segments coding for short bridge peptides were constructed and expressed in Escherichia coli. Surprisingly, the chimeric genes complemented the strain in which rbsA and rbsC were deleted. The chimeric proteins were overproduced, and the products were purified by affinity chromatography. In order to obtain highly purified protein, a poly-His leader peptide was incorporated so that Ni-chelate affinity chromatography could be employed. The leader peptide and the bridge peptide were designed with factor Xa-cleavable sites to permit recovery of the individual RbsA and RbsC protein. A rbsC gene encoding a poly-His leader was also constructed and expressed. Both the chimeric RbsA-C species and the poly-HisRbsC were produced at levels that permitted isolation of the equivalent of milligram quantities of RbsC per liter of culture. This is a substantial increase in amounts from any previous RbsC production vectors. All proteins from the rbs operon have now been overproduced and substantially purified.     

矩阵的加号逆理论在参数估计中的应用

本文从矩阵的加号逆理论出发,根据求矛盾线性方程组最佳逼近解的方法,建立起一种新的参数估计方法,同时给出了显著性检验方法,这种方法更简单更精确．     

一种具有纤溶活性的枯草杆菌蛋白激酶的研究──Ⅰ高产酶菌株的筛选与鉴定

本文主要阐述了一种具有纤溶活性的枯草杆菌（Ｂａｃｉｌｌｕｓｓｕｂｔｉｌｉｓ）蛋白激酶产生菌株的筛选与鉴定的研究结果。作者从初筛的１２株Ｂａｃｉｌｌｕｓｓｕｂｌｉｌｉｓ菌中,通过对固体发酵和液体发酵所产生的枯草杆菌蛋白激酶,用琼脂糖－纤维蛋白平板法测其活性,经比较不同菌株的活性,筛选出两株高产酶菌株：Ｂ．ｓｕｂｔｉｌｉｓＨＷ—１２和Ｂ．ｓｕｂｔｉｌｉｓＨＷ—３。同时对菌体和菌落形态特点、生理生化反应进行了鉴定,认为Ｂ．ＳｕｂｔｉｌｉｓＨＷ－１２菌株可用来做为发酵生产该酶的菌种。     

人胸腺素α原cDNA的克隆及在大肠杆菌中的表达

采用基因工程方法制取人胸腺素α原获得成功。用２０ｕｇ／ｍｌ植物血球凝集素（ＰＨＡ）和５００Ｕ／ｍｌ重组人白细胞介素２（ＩＬ－２）联合刺激人胎儿胸腺细胞,从中提取总ＲＮＡ,经反转录ＰＣＲ获得了人胸腺素α原ｃＤＮＡ;将之克隆入ｐＵＣ１９中,序列测定表明与已报道序列一致,进一步将之亚克隆入原核表达载体ｐＢＶ２２０,转化大肠杆菌ＤＨ５ａ．观察到在不改变氨基酸编码的前提下,增加胸腺素ａ原上游引物中Ａ、Ｔ含量,可以明显提高胸腺素α原的表达量,同时,不同培养基对它的表达也有影响。胸腺素α原在大肠杆菌中以可溶形式表达,不需复性。初步活性测定显示,它可明显刺激人外周血淋巴细胞Ｅ－玫瑰花结形成率。重组人胸腺素α原在大肠杆菌中表达,为其临床应用及基础研究奠定了基础。