节肢动物的基因组图和基因组学

W．Fiers等人1972年首次破译基因序列，开辟了遗传学的新时代，即基因组学时代。此后，很多生物的基因序列被破译。这些里程碑式的发现推动了生命科学的发展。20世纪的最后20年里，出现了聚合酶链式反应及克隆技术。基因组图实验、克隆基因、转录等过程中的大量数据需要分析储存，     

A型肉毒杆菌毒素的基因测序与分析

对国内A型肉毒杆菌毒素基因测序,分析了解毒素基因结构并推测其氨基酸序列。提取肉毒杆菌总DNA,利用自行设计的引物对目的基因进行PCR扩增,把目的基因导入E.Coli JM109,筛选含目的基因的阳性克隆菌进行测序和分析,从而了解毒素基因结构并推测其氮基酸序列。对A型肉毒杆菌毒素基因的核苷酸序列成功测序并与GenBank中的相应序列比较,其序列同源性为96％。推测其氨基酸序列同源性为100％。成功地对A型肉毒杆菌毒素的基因进行了测序,这为肉毒杆菌毒素中毒的快速诊断,以及进一步探求以基因工程方法生产此毒素奠定基础。     

Anchored PCR （A-PCR）： A new method for chromosome walking

PCR-based techniques are most popular methods for isolation of DNA sequences flanking a known region. Such techniques published to date mainly include three types: inverse PCR (IPCR), ligation-mediated PCR (LM-PCR) and randomly primed PCR (RP-PCR). IPCR was the first method developed for this kind of purpose. However, it is now rarely used because of the difficulty in finding suitable restriction sites in the target region or poor circularization of the template molecule. LM-PCR and RP-PCR are more frequently used nowadays, yet they also have some limitations, For example.     

Asian origin for Polystichum （Dryopteridaceae） based on rbcL sequences

Chloroplast rbcL sequences of 60 species of Polystichum sensu lato (s.l.), including 23 new sequences from southwest China, were used to assess the phylogenetic relationships within the genus. On the basis of estimated evolution rate of rbcL gene and the genetic distance data that passed relative-rate tests, we further estimated the divergence times between some clades of the genus. The phylogenetic relationships were inferred using the neighbor-joining and maximum-parsimony methods, both methods producing trees with completely congruent topology. These trees reveal that all species of Polystichum s.l. in this study (including Cyrtomium and Cyrtomidictyum) form a monophyletic group. The basal split in Polystichum s.l. separates a clade with all Asian members from a clade containing other species from all over the world. The phylogenetic and divergence time estimation results lead us to suggest that Polystichum s.l. originated in Asia in the late Late Cretacous (≈76 Ma) and migrated into other places in the world in early Eocene(≈46 Ma).     

Construction and characterization of a normalized whole-life-cycle cDNA library of rice

A cDNA library with genomic complete coverage is a powerful tool for functional genomic studies.For studying the functions of rice genes on a large scale,a normalized whole-life-cycle cDNA library is constructed based on the strategy of saturation hybridization with genomic DNA using rice cultivar Minghui 63,an elite restorer line for a number of rice hybrids that are widely cultivated in China,This library consists of cDNA from 15 directionally cloned cDNA libraries constructed with different tissues from 9 developmental stages.For normalization,the denatured plasmids purified from the 15 directionally cloned libraries are mixed and hybridized with saturated genomic DNA labeled with magnetic beads in two complementary systems. Well-matched plasmids are captured from the hybridized genomic DNA and electroporated into competent DH10B E. coli for construction of the normalized whole-life-cycle cDNA library.This library consists of 62000 clones with an average insert length about 1.4kb.Inverse Northern blotting shows that this cDNA library included many rarely expressed genes and tissue-specific genes.Sequencing of 10750 cDNA clones of this library reveals 6399 unique EST s(expressed sequence tags),indicating that the non-redundancy of the library is about 59.5%.This library has been used to make cDNA microarrays for functional genomic studies.     

Studies of new EST-SSRs derived from Gossypium barbadense

Existing cotton EST-SSR markers are mostly derived from Gossypium arboreum and Gossypium hir-sutum, but EST-SSR markers from Gossypium barbadense are scarce. One hundred and nineteen EST-SSRs were developed based on 98 unique ESTs from a cDNA library constructed in our laboratory using developing fibers from G. barbadense cv. Pima3-79. Among the SSRs, trinucleotide AAG appeared at a high frequency of 11.76%. 36 accessions (consisting of 13 diploids of the A genome, 11 diploids of the D genome and 12 allotetraploids of the AD genome) were employed to test new EST-SSRs. 76 EST-SSRs were successfully amplified, and 313 polymorphic fragments were yielded, with an average of 4.11 fragments per primer pair. The PIC ranged from 0.17 to 0.95 with an average of 0.53. Based on Jaccard’s genetic similarity coefficient, these 36 accessions were clustered into three groups. 21 EST-SSRs exhibited polymorphisms in BC1 population ((Emian22 × Pima3-79) × Emian22), 24 polymor- phic loci were generated, while 22 of the 24 polymorphic loci were integrated with our interspecific BC1 backbone genetic linkage map, and anchored in 12 chromosomes. This study effectively proved that EST-SSRs from G. barbadense are valuable for genetic diversity analysis and genetic mapping.     

Prediction and systematic study of protein-protein interaction networks of Leptospira interrogans

Leptospirosis, a serious and contagious zoonotic bacterial disease that havocs livers and kidneys in hu-mans and some animals, occurs in both urban and rural environments worldwide. It is caused by the genus lep- tospira, a corkscrew-shaped bacterium[1]. Though the genomes of Leptospira interrogans serovars Lai strain lai[2] and Leptospira interrogans serovars Copenhageni strain L1-130[3] were recently sequenced and many molecular and cellular studies on leptospires have been conducted, the …     

一株黄原胶降解菌的筛选与鉴定

从土壤中分离出一株能够降解黄原胶的菌株,对这株黄原胶降解菌进行形态特征、生理生化特征及16SrDNA的序列分析,并利用系统发育分析等研究结果,确认该菌株属于芽胞杆菌科的芽胞杆菌属,为进一步开展菌株改良、定点诱变等提高菌株降解黄原胶能力的工作奠定基础．     

新疆绵羊β-乳球蛋白(BLG)5′端调控区的克隆与序列测定

用聚合酶链式反应(PCR)技术,从新疆绵羊全血中扩增了β-乳球蛋白5′端调控区890bp的片段.通过T4 DNA连接酶将其连接于质粒载体pGEM-T上,转化至受体菌DH5α中.提取质粒经PCR扩增、酶切鉴定,证明重组质粒pT-BLG中含有β-乳球蛋白5′端调控区基因片段.经核苷酸序列测定,克隆的片段与发表的基因序列高度一致.     

Phylogenetic position of three Condylostoma species （Protozoa,Ciliophora, Heterotrichea） inferred from the small subunit rRNA gene sequence

The systematically poorly known ciliate genus Condylostoma was erected by Vincent in 1826. About 10 morphotypes have been reported, but any molecular investigations concerning this group so far are lacking. In this work, the small subunit ribosomal RNA （SS rRNA） gene of three marine Condylostoma species was sequenced, by which the phylogenetic trees were constructed by distancematrix, maximum parsimony and Bayesian inference methods. The results show that （1） all the trees have similar topologies with high supports; （2） Condylostoma is mostly related to the genus Condylostentor; and （3） three Condylostoma species as well as Condylostentor auriculatus cluster together and form a sister group with other heterotrichs. This is moderately consistent with the assessment of phylogenetic relationships of Condylostoma-related heterotrichs from the morphological information. The phylogenetic relationship of some other related heterotrichs, Peritromus, Folliculina, Stentor and Blepharisma, has been also discussed.