Sequence of an oleocin cDNA from Brassica napus

Antibodies raised against purified rapeseed 19 kDa oleosin protein were used to screen an embryo-derived gt11 expression library from Brassica napus. A near full-length cDNA clone, BnV, was isolated. The 781 bp cDNA contained an open reading frame of 549 bp followed by an untranslated region of 222 pb and a poly(A) region of 10 bp. Comparisons between this cDNA and a different oleosin cDNA previously isolated from the same library showed high degrees of sequence similarity in the central domain region and in the 3 untranslated region. Sequence similarities between the derived protein sequence of this cDNA and all other known oleosin protein sequences are discussed.     

Comparative Studies of the Seed Proteins of Brassica campestris, Brassica oleracea, and Brassica nigra

The albumin and globulin fractions of the seeds of Brassicacampestris, B. oleracea, and B. nigra have been investigatedusing serological methods and separation by acrylamide gel electrophoresisin order to determine the usefulness of these techniques fortaxonomic studies of the genus Brassica. The results of theprotein analysis given in this paper agree with establishedtaxonomy and suggest that both methods may prove useful in varioustaxonomic problems in the genus Brassica.     

An Antifungal Protein Purified from Pearl Millet Seeds Shows Sequence Homology to Lipid Transfer Proteins

In the course of a search for antifungal proteins from plant seeds, we observed inhibition of mycelial growth of Trichoderma viride with extracts of pearl millet. We have identified several proteins with antifungal properties in the seeds of pearl millet. One of these proteins has been purified to homogeneity and characterized. The purified protein has a molecular mass of 25 kDa. The N-terminal sequence of the protein (25 residues) shows homology to non-specific lipid transfer proteins (LTPs) of cotton, wheat and barley. The purified LTP inhibited mycelial growth of T. viride and the rice sheath blight fungus, Rhizoctonia solani in vitro.     

cDNA cloning and characterization of enolase from Chinese cabbage, Brassica campestris ssp. Pekinensis.

An enolase-encoding cDNA clone from Chinese cabbage, Brassica campestris ssp. Pekinensis, was isolated. This gene (Accession number: AY307448) had a total length of 1580bp with an open reading frame of 1335bp, and encoded a predicted polypeptide of 444 amino acids with a molecular weight of 47.38 kDa. The deduced amino acid (aa) sequence shared identity with a number of enolases ranging from Bacillus subtilis to human beings and had much higher identity with other plant enolases than with enolases from Bacillus, yeast and human beings. Comparison of its primary structure with those of other enolases revealed the presence of an insertion of 5 amino acids in enolase of Chinese cabbage. Expression of the cloned enolase gene decreased under salt stress, but increased in response to low temperature. Southern blot analysis of genomic DNA indicated that low-copies of enolase gene were present in the genome of Chinese cabbage.     

芸苔（菜心）茎叶中的几丁质结合蛋白的分离纯化和某些特性分析

用脱氨再生几丁质亲和层析和羧甲基-纤维素CM52离子交换柱层析从芸苔（菜心）的茎叶中纯化了4种几丁质结合蛋白：CBPa,CBPb、CBPc和CBPd,SDS-PAGE显示CBPa和CBPb的分子量分别为29．04kDa和30．68kDa。凝胶过滤方法测定CBPa和CBPb的分子量分别为29．04kDa和31．2kDa,表明两者都是单体酶。4种CBP蛋白均有几丁质酶活性,其中CBPa和CBPb还有溶菌酶活性。EDTA对CBPa和CBPb溶菌酶的活性均无影响,各种类型的几丁质对CBPa和CBPb都有较强的吸附作用,PAS染色法分析表明CBPa和CBPb均为糖蛋白。     

Purification and Partial Characterization of Arginine Decarboxylase from Brassica campestris

Arginine decarboxylase (EC 4.1.1.19) has been purified and characterized from Brassica campestris cv B-9. The enzyme was purified 1120 fold and the recovery was 9%. The mol wt of the enzyme determined by gel filtration was 240 kD with identical subunits of 60 kD. The pH and temperature optima for the enzyme were 8.0 and 30°C respectively. The Km was 0.31mM. Polyamines inhibited the enzyme activity significantly. Immunodiffusion with ADC-specific antibodies showed cross reactivity against purified ADC from Brassica.     

Exploring Sequence Characteristics Related to High-Level Production of Secreted Proteins in Aspergillus niger

Protein sequence features are explored in relation to the production of over-expressed extracellular proteins by fungi. Knowledge on features influencing protein production and secretion could be employed to improve enzyme production levels in industrial bioprocesses via protein engineering. A large set, over 600 homologous and nearly 2,000 heterologous fungal genes, were overexpressed in Aspergillus niger using a standardized expression cassette and scored for high versus no production. Subsequently, sequence-based machine learning techniques were applied for identifying relevant DNA and protein sequence features. The amino-acid composition of the protein sequence was found to be most predictive and interpretation revealed that, for both homologous and heterologous gene expression, the same features are important: tyrosine and asparagine composition was found to have a positive correlation with high-level production, whereas for unsuccessful production, contributions were found for methionine and lysine composition. The predictor is available online at http://bioinformatics.tudelft.nl/hipsec. Subsequent work aims at validating these findings by protein engineering as a method for increasing expression levels per gene copy.     

油菜叶绿体基因ndhB的序列

1　Ｓｏｕｒｃｅ　ＴｈｅｓｅｑｕｅｎｃｅｗａｓｄｅｔｅｒｍｉｎｅｄｆｒｏｍａＰＣＲ ｐｒｏｄｕｃｔ,ｗｈｉｃｈｗａｓｌｉｇａｔｅｄｔｏｐＢｌｕｅｓｃｒｉｐｔＳＫ( )ｖｅｃｔｅｒ (ｎａｍｅｄａｓｐＳＮ) ,ｆｒｏｍｔｈｅｃｈｌｏｒｏｐｌａｓｔｇｅｎｏｍｉｃＤＮＡｏｆＢｒａｓｓｉｃａｎａｐｕｓ(Ｏｉｌｓｅｅｄｒａｐｅ)ｃｖ.Ｈ1 65.2　Ｎａｍｅａｎｄｄｅｓｃｒｉｐｔｉｏｎ　Ｂｒａｓｓｉｃａｎａｐｕｓｃｈｌｏｒｏ ｐｌａｓｔｇｅｎｅｎｄｈＢｉｓ 2 4 67ｂｐｌｏｎｇｗｉｔｈｔｗｏｅｘｏｎｓｔｈａｔｅｎｃｏｄｅ 51 0ａ…     

油菜叶片硝酸还原酶同功酶的纯化与特性

通过对硝酸还原酶（ＮＲ）亲和层析洗脱过程的部分改进,从油菜叶片中分离纯化到诱导型（ｉＮＲ）及组成型（ｃＮＲ）两种硝酸还原酶同功酶。电泳分析表明两者均达到银染单带纯。ｃＮＲ分子量（ＭＷ）为４５０ｋＤ．ｉＮＲＭＷ为２２０ｋＤ;两者亚基ＭＷ均为１１０ｋＤ,但亚基数目不一样,氨基酸组成也有差异。ｉＮＲ与ｃＮＤ的等电点ｐＨ不同,分别为４．４及６．０,免疫交叉反应显示ｃＮＲ的抗原性为ｉＮＲ的７５％。     

Cell wall degrading enzymes in Orobanche aegyptiaca and its host Brassica campestris

Activities of the cell-wall degrading enzymes cellulase (EC 3. 2. 1. 4). poly-galacturonase (EC 3. 2. 1. 15) and xylanase (EC 3. 2. 1. 8), and also that of protease (EC 3. 4. 21. 14) have been determined in the parasite Orobanche aegyiptiaca Pers. and in the roots of infected and control host plants of Brassica campestris L. var. rapa Hartm. Shoots, roots and root-freed tubers of Orobanche showed pronounced activities of these enzymes. When expressed in terms of specific activity, protease, cellulase and polygalacturonase activities were higher and xylanase lower in all parts of Orobanche than in the roots of infected or control host plants. Orobanche roots had higher activities of cellulase, polygalacturonase and protease than tuber or shoot. While xylanase activity was higher in the shoot than in roots or tuber. Infection by Orobanche significantly enhanced the cellulase activity of host roots. Cellulase and polygalacturonase in the Orobanche roots appear as the main enzymes involved in establishing haustorial connection with the host root. Protease might be involved in degrading proteins and lipoproteins of the cell walls and membranes of the host tissues.     

不结球白菜晚抽薹BcFLC3基因的克隆及表达分析

根据大白菜BcFLC3基因(GenBank登录号AY036890.1)保守域序列设计引物,扩增不结球白菜晚抽薹BcFLC3基因的核心片段,结合RACE技术获得该基因1 017 bp的全长cDNA序列.序列分析结果表明,该cDNA包含完整开放阅读框,编码197个氨基酸的蛋白质,其分子量为21.62 kD,等电点9.36.荧光定量 PCR分析表明,不结球白菜经4℃低温处理后,BcFLC3基因在叶片中的表达量抽薹前明显高于抽薹后;低温处理后BcFLC3基因在不同部位的表达量存在明显差异,表达量由高到低依次为叶、茎、花蕾、花和根.Southern杂交结果表明,BcFLC3基因在不结球白菜基因组中为多拷贝.     

Sequence and molecular analysis of the rpoA cluster genes from Xanthomonas campestris pv. campestris

The Xanthomonas campestris rpsM (S13)-rpsK (S11)-rpsD (S4)-rpoA (alpha)-rplQ (L17) cluster, encoding RNA polymerase alpha-subunit and four ribosomal proteins, reside in a 3164-bp DNA region. The N-terminal sequence of the authentic alpha-protein determined chemically matches that predicted from the nucleotide sequence. rplQ is monocistronic, instead of being co-transcribed with the other genes as in Escherichia coli. Antiserum against the His-tagged alpha-protein cross-reacted with the E. coli alpha-protein.     

Sequence and structure of Brassica rapa chromosome A3

Background

The species Brassica rapa includes important vegetable and oil crops. It also serves as an excellent model system to study polyploidy-related genome evolution because of its paleohexaploid ancestry and its close evolutionary relationships with Arabidopsis thaliana and other Brassica species with larger genomes. Therefore, its genome sequence will be used to accelerate both basic research on genome evolution and applied research across the cultivated Brassica species.

Results

We have determined and analyzed the sequence of B. rapa chromosome A3. We obtained 31.9 Mb of sequences, organized into nine contigs, which incorporated 348 overlapping BAC clones. Annotation revealed 7,058 protein-coding genes, with an average gene density of 4.6 kb per gene. Analysis of chromosome collinearity with the A. thaliana genome identified conserved synteny blocks encompassing the whole of the B. rapa chromosome A3 and sections of four A. thaliana chromosomes. The frequency of tandem duplication of genes differed between the conserved genome segments in B. rapa and A. thaliana, indicating differential rates of occurrence/retention of such duplicate copies of genes. Analysis of 'ancestral karyotype' genome building blocks enabled the development of a hypothetical model for the derivation of the B. rapa chromosome A3.

Conclusions

We report the near-complete chromosome sequence from a dicotyledonous crop species. This provides an example of the complexity of genome evolution following polyploidy. The high degree of contiguity afforded by the clone-by-clone approach provides a benchmark for the performance of whole genome shotgun approaches presently being applied in B. rapa and other species with complex genomes.