Cloning and expressional analyses of a cinnamoyl CoA reductase cDNA from rice seedlings

Cinnamoyl CoA reductase (CCR: EC 1.2.1.44),the entry-point enzyme of the llgnin specific biosynthetic pathway, catalyzes the conversion of cinnamoyl CoA esters to their corresponding dnnamaldehydes. Multiple sequence alignment showed that the deduced polypeptide shared 70% similarity and 30% sequence identity at the amino acid level with defined CCR genes from other plant species and they all contain the common signature sequences thought to be the catalytic site as well as the putative NADP binding domain.Using a conserved OsCCR cDNA fragment as the probe for library screening, we isolated the genomic DNA that covered the whole coding region of OsCCR with total length of 3045bp including 4 introns and 5 exons. The open reading frame for our OsCCR gene coBtAin~ 337 amino adds. Northern blot indicated that OsCCR was expressed in different organs with the highest level found in stems. In situ hybridization results showed that OsCCR mRNA was localized mainly along the vascular bundles in stems and leaves, and also in lateral roots that was differentiating from the tiilering node. We conclude that the vascular-localized expression of OsCCR gene may suggest its possible involvement in llgnin biosynthesis. Cloning and characterization of OsCCR will help to clarify how llgniflcations in plants are regulated and will provide a physical basis for creating genetically engineered rice plants with optimal lignin contents.     

Importance of NPA motifs in the expression and function of water channel aquaporin-1

The asparagine-proline-alanine sequences (NPA motifs) are highly conserved in aquaporin water channel family. Crystallographic studies of AQP1 structure demonstrated that the two NPA motifs are in the narrow central constriction of the channel, serving to bind water molecules for selective and effi-cient water passage. To investigate the importance of the two NPA motifs in the structure, function and biogenesis of aquaporin water channels, we generated AQP1 mutations with NPA1 deletion, NPA2 de-letion and NPA1,2 double deletion. The coding sequences of the three mutated cDNAs were subcloned into the mammalian expression vector pcDNA3.1 to form expression plasmids. We established stably transfected CHO cell lines expressing these AQP1 mutants. Immunofluorescence indicated that all the three mutated AQP1 proteins are expressed normally on the plasma membrane of stably transfected CHO cells, suggesting that deletion of NPA motifs does not influence the expression and intracellular processing of AQP1. Functional analysis demonstrated that NPA1 or NPA2 deletion reduced AQP1 water permeability by 49.6% and 46.7%, respectively, while NPA1,2 double deletion had little effect on AQP1 water permeability. These results provide evidence that NPA motifs are important for water per-meation but not essential for the expression, intracellular processing and the basic structure of AQP1 water channel.     

植物转化酶及其基因表达与调控

蔗糖利用依赖于蔗糖的降解.植物转化酶通过降解蔗糖为己糖,不仅为呼吸作用提供底物,而且作为信号分子调节多种基因的表达,同时转化酶基因的表达也受到多种因素的调控.综述了近年来植物转化酶的种类、特性、功能及其基因表达在转录水平和翻译水平调控机制的研究进展.     

TaNHX2基因植物表达载体的构建及在烟草中的表达

将TaNHX2基因重组于质粒pBIN438的CaMV 35S启动子下游,构建含TaNHX2基因的植物双元表达载体pBIN438-TaNHX2.采用农杆菌介导转化烟草(Nicotiana tobacum L.),获得含TaNHX2的转基因烟草植株.经PCR、RT-PCR分析表明,TaNHX2基因已整合到烟草中,并且得到表达.耐盐分析表明,外源基因TaNHX2提高了转基因植株的耐盐性.     

植物细胞膜H^＋-ATPase对必需矿质养分的适应性

植物细胞膜H^＋-ATPase是初级转运蛋白，在各种离子和代谢产物的跨膜运输中起着重要的作用；介绍了植物细胞H^＋-ATPase的结构、生化特性、生理功能与调控机制；重点综述了细胞膜H^＋-ATPase的活性及其蛋白数量与基因表达对植物必需矿质养分的适应性。     

聚离子亚基化合物介导的基因转移方法可用于外源基因？…

利用聚离子亚基化合物介导的基因转移方法，研究了外源β－半乳糖苷酶报告基因的瞬时表达。结果表明，聚离子亚基化合物介导的基因转移方法可用于外源基因的瞬时表达研究。而且在相同的条件下，用该方法在ＮＩＨ３Ｔ３细胞和ＰＡ３１７细胞中对报告基因转移的效率     

噬菌体展示外源多肽的免疫原性研究

利用噬菌体展示技术将丝状噬菌体fd基因8克隆入pKK223-3质粒中，将白色念珠菌特异表位基因插入到修饰后的质粒载体中，并制备了杂合噬菌体，利用纯化的该表位抗原PA免疫小鼠，结果表明该噬菌体-表位抗原PA在有无佐剂存在的情况下均产生抗体，用ELISA方法采用双波长（450/630nm)检测抗体，其在有无佐剂存在的情况下光密度值无明显差别，免疫后小鼠脾淋巴细胞在有无佐剂的情况下均产生明显增殖，未见明显的毒副作用，具有良好的安全性，该方法产生的抗体与传统的合成多肽相比具有方法简便、低价高效等特点，它将成为生产高效低价疫苗的有效途径，该噬菌体-表位抗原PA为真菌疫苗的研制打下基础。