大肠杆菌丝氨酸羟甲基转移酶基因(glyA)的克隆和表达

利用插入失活及营养缺陷型互补法将大肠杆菌K12 13kb的glyA基因克隆到质粒pBR329中。将重组质粒酶切,亚克隆,确定2.6kb PstI-EcoRI亚克隆片段带有完整的glyA基因。共获得12株glyA基因重组菌,对重组质粒进行了酶切鉴定。不同重组菌丝氨酸羟甲基转移酶(SHMT)活性及其酶表达量均不相同。受体菌未检测到丝氨酸的产生。重组菌株JM109(pSM13)、K12(pSM13)、K12(pSM14)和K12(pSM15)SHMT酶表达量分别占全菌可溶性蛋白的15.7％、15.4％、11.8％和9.48％。     

抗破伤风类毒素单抗可变区基因的克隆及在大肠杆菌中表达的三种工程抗体

The heavy and light chain variable region genes of anti-tetanus toxoid (TT) antibody and the heavy chain Fd genes were amplified and cloned through RT-PCR from mouse hybridoma cells. The sequences of VH and VK were determined. Fd gene fragments were expressed in E. coli. The ELISA results indicated that the expressed Fd showed antigen binding activity but was nonspecific. Furthermore, through SOE and PCR techniques, the VH and VK gene fragments together with ScFv linker were assembled into single chain antibody (ScFv) gene fragment. While together with human heavy chain CH 1 gene fragment and Fab linker, they were assembled into chimeric Fab gene fragment. The two assembled gene fragments were separately inserted into phagemid pHEN 1, which was a fd-based vector containing gene 3 encoding the minor coat protein. In presence of helper phage M 13-VCS the anti-TT phage-ScFv or phage-Fab were displayed on the surface of phage particles respectively. Results from phage-ELISA indicated that both phage antibodies were TT-specific.     

在大鼠发育过程中β1,4半乳糖基转移酶的组织分布及其变化的研究

用HPLC纯化了荧光标记的底物（Gnβ1-2Ma1-6（Gnβ1-2Mα1-3）Mβ1-4Goβ1-4Gn-PA）用β-1,4-半乳糖基转移酶的荧光标记底物的HPLC测定方法,测定了在发育过程中大鼠肝,肾,脑中的酶活性的变化,结果表明,（1）在正常成年大鼠中,各组织酶活性具有组织特异性,（2）不同的发育期,其酶活性不同。胎鼠（孕20d,以下同）时最高,以后就渐渐下降,各组织酶活力变化幅度是不一致的,这些变化的生理意义有待子进一步研究．     

香菇空间诱变突变体的分子生物学鉴定研究*

香菇Cr04菌株经空间诱变后从中筛选出了农艺性状明显改善的突变菌株Cr04DZ。本研究用RAPD和AFLP技术对Cr04DZ与其地面对照菌株Cr04进行了DNA指纹分析,找出了它们之间的DNA多态性,从而从分子水平上证实了香菇经空间诱变后遗传物质发生了变化。此外,还比较了AFLP与RAPD检测香菇DNA多态性的效率,优化了对食用菌进行RAPD与AFLP分析的反应条件。通过对Cr04DZ及其对照株Cr04的RAPD及AFLP分析,筛选出了突变体的3个RAPD及29个AFLP多态性产物,已完成了3个AFLP产物的克隆。     

Construction and characterization of a bacterial artificial chromosome library of thermo-sensitive genie male-sterile rice 5460S

In order to develop a detailed physical map of the thermo-sensitive genie male-sterile (TGMS) gene-encompassing region and finally clone the TGMS gene, a high-quality rice bacterial artificial chromosome (BAC) library from TGMS rice 5460S was constructed. The method of constructing BAC library was examined and optimized. The 5460S library consists of 19 584 BAC clones with an average insert size of 110 kb, which represents about 5 times rice haploid genome equivalents. Rice inserts of up to 140 kb and 250 kb were isolated and appeared stable after 100 generations of serial growth. Hybridization of BAC clones with mitochondrial and chloroplastic genes as probes demonstrated that this library has no organellar contamination. The 5460S library was screened with 3 molecular markers linked to tmsl gene as probes and at least 1 BAC clone was identified with each probe. The insert ends of positive clones were successfully isolated using thermal asymmetric interlaced PCR (TAIL-PCR) technique.     

光活性除草剂的研制

<正> 美国厄巴纳香潘的伊利诺伊大学Coustautiu Rebeiz和hetbett Hopen为首的一个小组研制出一种新的能高效地杀死杂草的光活性除草剂。这种除草剂的主要成份是氨基乙酰丙酸(ALA),这是一种正常存在于动植物体内的氨基酸。夜间喷雾,ALA被杂草吸收并迅速代谢成为对光非常敏感的四吡咯,在阳光下四吡咯正常地形成叶绿素。但是,在暗处,由ALA获得的四吡咯可以积累到反常的高水平。随着太阳光的出现,它们开始参与能大量产生杀死     

日本揭示了生物技术的趋势和预测

<正> 到2000年,日本生物技术在工业上的应用预计可创造5140亿美元收入,约占国民生产总值的10%。其中生物工业的传统技术占4543亿美元,新生物技术大约占607亿。该预测是日本工业发酵协会生物工业开发中心汇编的一份调查报告中的一部分。鉴于这种预测,人们对1985年日本政府6个机构在生物技术研究与发展方面花费13460万美元的数额(超出1984年     

从原生质体再生水稻

<正> 据国家农业生物资源研究所宣称,从原生质体已首次再生出水稻植株。该所(农林水产省的一个研究单位)的研究人员分别培养出60种原生质体。其中中,有4种能生根和发芽,有一种可长成10厘米高的完整植株。20种原生质体来自种子细胞,而其它40种是由某些花粉细胞中分离的细胞壁,这些花粉细胞是从单个花粉细胞繁殖来的。     

Amersham公司出售Amgen公司生产的生长因子

<正> 从1月初开始,Amersham国际公司将一些遗传操作产生的生长因子包括干扰素和间白细胞素(T-细胞生长因子)列入供科学研究用的产品中。这些产品是由Amersham公司用重组DNA技术制造的第一批产品中的一部分。这些生长因子是由索仁奥克斯Amgen