乙型肝炎表面抗原S—蛋白糖化位点序列的人工变异及其哺乳动物细胞表…

利用末端重叠ＰＣＲ法和定点突变技术，获得了去除Ｎ－连接多糖结构的乙肝表面抗原Ｓ突变基因，将乙肝表面抗原Ｓ－蛋白中结合Ｎ－连多糖结构序列Ａｓｎ－Ｘ－Ｔｈｒ中的第１４８苏氨酸的密码子ＡＣＴ，改变为编码甘氨酸的密码子ＧＧＴ。构建了该突变基因的表达载体并在ＣＨＯ细胞中表达，筛选到两株表达水平较高的细胞系。对其中的一株细胞系Ｃ４１的表达产物做了初步纯化和鉴定，纯化样品经ＳＤＳ－ＰＡＧＥ电泳分析，只含有２３ｋ     

番茄rin位点上的一个MADS盒基因调控果实成熟

ｒｉｎ番茄突变体的乙烯生成和果实成熟均受到抑制 ,除此之外还表现萼片增大和花序决定性丧失。尽管该突变体维持良好的乙烯敏感性 ,但果实成熟却不能被外源乙烯诱导 ,这暗示ＲＩＮ基因编码的产物同时调控依赖于和不依赖于乙烯的成熟过程。作者先前的研究已表明ｒｉｎ位点定位于番茄第 5染色体。本文报道通过定位克隆发现ｒｉｎ位点上存在两个串联的ＭＡＤＳ盒基因 ,分别定名为ＬｅＭＡＤＳ＿ＲＩＮ和ＬｅＭＡＤＳ＿ＭＣ ,前者调控果实成熟过程而后者则影响萼片先育和花序决定性。这两个基因的功能通过正义转化ｒｉｎ番茄和反义转化野生型番…     

噬菌体整合酶介导的位点特异性基因治疗

将治疗基因整合到患染色体上的特定位点并实现稳定的表达是基因治疗的关键,包括病毒载体和转座子系统在内的几种技术已经用于整合,但都存在治疗基因负载力较小及随机整合的应用障碍。来自链霉菌噬菌体中C31、R4及来自乳酸乳球菌噬菌体TP901-1的整合酶,能够识别有限数量的天然基因组序列(假att位点)．从而催化治疗基因位点特异性地整合进入高等真核生物基因组。这种新型的噬菌体整合酶系统在基因负载力及染色体的特异性整合方面具有很大的优越性。迄今为止,这种系统已经成功用于几种基因治疗的研究模型中．将会成为基因治疗的有用工具。     

吡咯赖氨酸：第 22 种参与蛋白质生物合成的氨基酸

吡咯赖氨酸在产甲烷菌的甲胺甲基转移酶中发现,是目前已知的第 22 种参与蛋白质生物合成的氨基酸,与标准氨基酸不同的是,它由终止密码子 UAG 的有义编码形成 . 与之对应的在产甲烷菌中也含有特异的吡咯赖氨酰 -tRNA 合成酶 (PylRS) 和吡咯赖氨酸 tRNA (tRNA^Pyl). tRNA^Pyl具有不同于经典 tRNA 的特殊结构 . 产甲烷菌通过直接途径和间接途径这两种途径生成吡咯赖氨酰 -tRNA^Pyl(Pyl-tRNA^Pyl) ,它还可能通过 mRNA 上的特殊结构以及其他还未发现的机制,控制 UAG 编码成为终止密码子或者吡咯赖氨酸 . 比较了吡咯赖氨酸与另一种非标准氨基酸,第 21 种氨基酸———硒代半胱氨酸的相似点与不同点 .     

等位基因特异性引物PCR技术及其应用研究

目的：研究建立等位基因特异性引物PCR技术体系,并将其应用于基因单核苷酸多态性研究工作。方法：通过美国国家生物信息中心(NCBI)的genBank获取基因序列及其相应位点的SNP信息。利用Primer5．0软件设计引物,并经NCBI的Blast2．0软件检验其特异性。结果：建立了单一等位基因特异性引物PCR(SASP—PCR)与嵌套式等位基因特异性引物PCR(NASP-PCR)两种技术,并应用于β2肾上腺素受体及内皮源性一氧化氮合酶基因单核苷酸多态性的研究,证实该技术的稳定性和优越性。结论：等位基因特异性引物PCR技术是一种更为简便、特异性较高、费用少的、便于推广的SNP检测方法,特别是在群体基因单核苷酸多态性研究中更有优势。     

基于支持向量机的人类5’非翻译区剪接位点识别

基因非编码区域剪接位点的识别是基因识别中一个非常具有挑战性的问题,尤其是5’非翻译区中剪接位点的识别。与一般剪接位点不同,5’非翻译区剪接位点的两侧不存在由编码到非编码的状态转移,所以通常的剪接位点识别算法在非翻译区的性能不太理想。文章采用了基于支持向量机的方法对5’非翻译区中的剪接位点进行识别。为了提高识别精度,采用了基于矩阵相似性度量的核函数参数选取方法,它能够简单快速地确定合适的核函数参数,进而提高核函数的识别性能。通过实验验证,经过参数选择后的支持向量机能够较好地识别5＇非翻译区剪接位点。     

中国人胃癌染色体17q21区域基因遗传不稳定性研究

To investigate microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396, D17S579 and D17S855, and their effect on the expression of nm23H1 and BRCA1 of gastric cancer, which would provide experimental basis for clinical treatment and prognosis analysis of gastric cancer. DNA was extracted from paraffin-embedded materials. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze MSI and LOH. Expression of nm23H, and BRCA1 was detected by Envision immuno-histochemistry and Leica-Qwin computer imaging techniques. In the forty cases of gastric cancer, the frequency of MSI, LOH and nm23H1 protein were 20.00%, 17.50% and 55.00% respectively at locus D17S396, while at locus D17S579, the frequency of MSI, LOH and BRCA1 protein were 22.50%, 15.00% and 37.50% respectively; at locus D17S855, the frequency of MSI, LOH and BRCA1 of thirty-seven cases were 18.92%, 18.92%, 37.84% respectively. In tumor node metastasis (TNM) staging, at locus D17S396, D17S579 and D17S855, MSI in stages I + II appeared more frequently than that in stages III + IV, while LOH appeared the contrary tendency. In the group of metastasis of gastric cancer, MSI had a less frequency (5.00%) than that with no metastasis (35.00%, P < 0.05) at locus D17S396, but LOH appeared more frequently (30.00%) than that with no metastasis (5.00%, P < 0.05). At locus D17S579, MSI had an increasing tendency with the degree of tumor differentiation (50.00% in high differentiation cases, 20.00% in middle differentiation cases, and 0% in low differentiation cases, P < 0.05). The frequency of nm23H1 and BRCA1 protein in stages TNM I + II was higher than that in stages TNM III + IV; and that in higher differentiation cases was higher than in poor differentiation cases. The frequency of nm23H1 protein in the group of metastasis (30.00%) was less than that with no metastasis significantly (80.00%, P<0.01). The frequency of nm23H1 protein in the group positive to MSI (87.50%) was higher than that in the group negative to MSI (46.88%, P < 0.05). However, nm23H1 protein in group positive to LOH (14.29%) was lower than that in the group negative to LOH (63.64%, P < 0.05). The frequency of BRCA1 protein in the group positive to MSI (66.67%) was more than that in the group negative to MSI (29.03%, P < 0.05). The results of experiments indicate that MSI and LOH may separately control the development of sporadic colon cancer with different pathways. MSI may be an early period molecule marker for sporadic colon cancer, enhanced expression of nm23H1 protein can effectively inhibit colon cancer metastasis and improve prognosis of sporadic colon cancer patients. By comparison, LOH mostly arises in the late period of sporadic colon cancer and endows a high aggressive and poor prognostic phenotype. nm23H1 protein could effectively restrain gastric cancer metastasis and development; and BRCA1 protein could restain tumor from becoming lower differentiation.     

细菌细胞分裂位点的调控机制及其研究进展*

大肠杆菌细胞内共有3个潜在的分裂位点,一个在细胞中部,另外两个位于细胞的两极。正常情况下,细菌仅利用中部的分裂位点以二分裂方式进行细胞的对称分裂。大肠杆菌细胞分裂时,中部潜在分裂位点的选择受到min操纵子(含minC、minD、minE3个基因)的精细调控。minC基因所编码的MinC蛋白是细胞分裂的抑制因子,与具有ATPase活性的MinD蛋白结合后被激活。在MinE蛋白的作用下,MinC和MinD蛋白在大肠杆菌细胞的两极间来回振荡。整个振荡周期中,MinC蛋白在细胞两极的两个潜在分裂位点处所停留的时间