Anti-infection peptidomics of amphibian skin

Peptidomics and genomics analyses were used to study an anti-infection array of peptides of amphibian skin. 372 cDNA sequences of antimicrobial peptides were characterized from a single individual skin of the frog Odorrana grahami that encode 107 novel an

大蹼铃蟾血管扩张因子及其制备方法和其基因

本发明涉及大蹼铃蟾血管扩张因子及其制备方法和其基因，属于生物医学领域。其血管扩张因子为从中国两栖类动物大蹼铃蟾(Bombinamaxima)皮肤分泌物中分离得到的一种单链多肽，分子量2985，等电点10.2，多肽氨基酸全序列一级结构为：NH2－GIGRKFLGGVKTTFRCGVKDFASKHLY－NH2。制备方法是收集大蹼铃蟾皮肤分泌物，离心去除沉淀、冷冻干燥后，经离子交换、高压液相反相柱层析分离纯化后即得到。编码血管扩张因子的基因由647个核苷酸组成，其中编码成熟血管扩张因子为第178－258位核苷酸。其血管扩张因子具有显著的扩张血管特别是微血管的作用，其基因作为基因工程制备血管扩张因子的应用。

Isolation and preliminary characterization of a 22-kDa protein with trypsin inhibitory activity from toad Bufo andrewsi skin

A novel trypsin inhibitor termed BATI was purified to homogeneity from the skin extracts of toad Bufo andrewsi by successive ion-exchange, gel-filtration and reverse-phase chromatography. BATI is basic single chain glycoprotein, with apparent molecular weight of 22 kDa in SDS-PAGE. BATI is a thermal stable competitive inhibitor and effectively inhibits trypsin's catalytic activity on peptide substrate with the inhibitor constant (K-i) value of 14 nM and shows no inhibitory effect on chymotrypsin, thrombin and elastase. The N-terminal sequence of BATI is EKDSITD, which shows no similarity with other known trypsin inhibitors. (c) 2005 Elsevier Ltd. All rights reserved.

Pontibacter akesuensis sp nov., isolated from a desert soil in China

A Gram-negative, non-motile, rod-shaped bacterium, designated strain AKS 1 T, was isolated from a desert soil sample collected from Alkesu, Xin.lang Province, China. A taxonomic study, including phylogenetic analysis based on 16S rRNA gene sequences and p

大蹼铃蟾抗微生物肽及其制备方法和其基因

本发明涉及一种大蹼铃蟾抗微生物肽及其制备方法和其基因，属于生物医学领域。抗微生物肽为从中国两栖类动物大蹼铃蟾(Bombina maxima)皮肤分泌物中分离得到的一种单链多肽，分子量2840，等电点983，多肽氨基酸全序列一级结构为：NH2－SIGAKILGGVKTFFKGALKELASTYLQ－NH2。制备方法是收集大蹼铃蟾皮肤分泌物，离心去除沉淀、冷冻干燥后，经离子交换、高压液相反相柱层析分离纯化后得到。编码抗微生物肽的基因由663个核苷酸组成，其中编码成熟抗微生物肽为第165－245位核苷酸。抗微生物肽具有广谱和显著的抑制细菌和真菌生长作用。抗微生物肽基因作为基因工程制备抗微生物肽的应用。

Maximin 9, a novel free thiol containing antimicrobial peptide with antimycoplasma activity from frog Bombina maxima

Amphibian skin is a rich resource of antimicrobial peptides, like maximins and maximin Hs from frog Bombina maxima. Novel cDNA clones encoding a precursor protein, which comprises a novel maximin peptide (maximin 9) and reported maximin H3, were isolated from two constructed skin cDNA libraries of B. maxima. The predicted primary structure of maximin 9 is GIGRKFLGGVKTTFRCGVKDFASKHLY-NH2. A surprising substitution is at position 16, with a free cysteine in maximin 9 rather than usual conserved glycine in other reported maximins. Maximin 9, the homodimer form and its Cys(16) to Gly(16) mutant were synthesized and their antimicrobial activities were evaluated. Unlike previously reported maximin 3, the tested bacterial and fungal strains were resistant to maximin 9, its homodimer and the Cys(16) to Gly(16) mutant (with MICs > 100 mu M). On the other hand, interestingly, while eight clinical Mollicutes strains were generally resistant to maximin 9 homodimer and its Cys(16) to Gly(16) mutant, most of them are sensitive to maximin 9 at a peptide concentration of 30 mu M, especially in the presence of dithiothreitol. These results indicate that the presence of a reactive Cys residue in maximin 9 is important for its antimycoplasma activity. The diversity of antimicrobial peptide cDNA structures encountered in B. maxima skin cDNA libraries and the antimicrobial specificity differences of the peptides may reflect well the species' adaptation to the unique microbial environments. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Multiple bombesin-like peptides with opposite functions from skin of Odorrana grahami

Bombesin-like peptides (BLPs) are a family of neuroendocrinic peptides that mediate a variety of biological activities. Three mature BLPs from the skin secretions of the frog Odorrana grahami were purified. Several bombesin-like peptide cDNA sequences enc

具有生物活性的纯化多肽及其制备的用于促进动物神经细胞生长、发育以及抑制其凋亡的药物

本发明为一种具有生物活性的纯化多肽及其制备的用于促进动物神经细胞生长、发育以及抑制其凋亡的药物，含有下列氨基酸序列(SEQ IDNO：1)，由适量的具有生物活性的纯化多肽和制药上可接受的赋形剂和稀释剂组成。主要用于促进动物神经细胞生长、发育，效果良好。

Identification and molecular cloning of a novel neuromedin U analog from the skin secretions of toad Bombina maxima

Amphibian skin contains rich neuropeptides. In the present study, a novel neuromedin U (NmU) analog was isolated from skin secretions of Chinese red belly Load Bombina maxima. Being 17-amino acids long, its primary structure was established as DSSGIVGRPFFLFRPRN-NH2, in which the C-terminal 8-residue segment (FFLFRPRN) is the same as that of rat NmU, while the N-terminal part DSSGIVGRP shows a great sequence variation compared with those of NmU peptides from different resources. The peptide, named Bm-NmU-17, was found to elicit concentration-dependent contractile effects on smooth muscle of rat uterus horns. The cDNA Structure of the peptide, as obtained by a 3'-RACE strategy and subsequently cloning from a skin cDNA library, was found to contain a coding region of 438 nucleotides. The encoded precursor is composed of 145 amino acids with a single copy of Bm-NmU-17 located towards the C-terminus. The sequence of the peptide is preceded by a dibasic site (Lys-Arg) and followed by the sequence of Gly-Arg-Lys, providing the sites of cleavage and releasing of the mature peptide. (c) 2005 Elsevier B.V. All rights reserved.

Bacillus pallidus sp nov., isolated from forest soil

A Gram-positive bacterium, designated strain CW 7(T), was isolated from forest soil in Anhui Province, south-east China. Cells were strictly aerobic, motile with peritrichous flagella and rod-shaped. The strain grew optimally at 30-37 degrees C and pH 7.0-8.0. The major fatty acids of strain CW 7(T) were anteiso-C-15:0, iso-C-15:0 and anteiso-C-17:0. The predominant menaquinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The G + C content of the genomic DNA was 42.3 mol%. Phylogenetic analysis indicated that strain CW 7(T) belonged to a monophyletic cluster within the genus Bacillus and showed 16S rRNA gene sequence similarities of less than 96.5% to recognized species of the genus Bacillus. The results of the polyphasic taxonomic study, including phenotypic, chemotaxonomic and phylogenetic analyses, showed that strain CW 7(T) represents a novel species of the genus Bacillus, for which the name Bacillus pallidus sp. nov. is proposed. The type strain is CW 7(T) (=KCTC 13200(T)=CCTCC AB 207188(T)=LMG 24451(T)).

酸性抗菌肽及其基因和在制药中的应用

本发明涉及一种酸性抗菌肽及其基因和在制药中的应用，属于生物医学领域。酸性抗菌肽，是中国两栖类动物大蹼铃蟾基因编码的一种单链多肽，分子量2021.4，等电点4.8，多肽氨基酸全序列一级结构为：Ile Leu Gly Pro Val LeuGly Leu Val Ser Asp Thr Leu Asp Asp Val Leu Gly Ile Leu-AMIDATION。编码酸性抗菌肽的基因由632个核苷酸组成，其中编码成熟酸性抗菌肽为第418-477位核苷酸。人工化学合成的酸性抗菌肽具有显著的抑制细菌生长的作用，可以作为制备抗微生物感染疾病药物的应用。

Bm-TFF2, a trefoil factor protein with platelet activation activity from frog Bombina maxima skin secretions

In mammals, trefoil factor family (TFF) proteins are involved in mucosal maintenance and repair, and they are also implicated in tumor suppression and cancer progression. A novel two domain TFF protein from frog Bombina maxima skin secretions (Bm-TFF2) has been purified and cloned. It activated human platelets in a dose-dependent manner and activation of integrin a(11b)beta(3) was involved. Aspirin and apyrase did not largely reduce platelet response to Bm-TFF2 (a 30% inhibition), indicating that the aggregation is not substantially dependent on ADP and thromboxane A2 autocrine feedback. Elimination of external Ca2+ with EGTA did not influence the platelet aggregation induced by Bm-TFF2, meanwhile a strong calcium signal (cytoplasmic Ca2+ release) was detected, suggesting that activation of phospholipase C (PLC) is involved. Subsequent immunoblotting revealed that, unlike in platelets activated by stejnulxin (a glycoprotein VI agonist), PLC gamma 2 was not phosphorylated in platelets activated by Bm-TFF2. FITC-labeled Bm-TFF2 bound to platelet membranes. Bm-TFF2 is the first TFF protein reported to possess human platelet activation activity. (c) 2005 Elsevier Inc. All rights reserved.

Molecular Phylogeny of Geographical Isolates of Bursaphelenchus xylophilus: Implications on the Origin and Spread of this Species in China and Worldwide

The genetic diversity and phylogeny of 26 isolates of Bursaphelenchus xlophilus from China, Japan, Portugal and North America were investigated based on the D2/3 domain of 28S rDNA, nuclear ribosomal Internal Transcribed Spacer (ITS) sequences, and random amplified polymorphic DNA (RAPD) analysis. The genetic diversity analysis showed that the D2/3 domain of 28S rDNA of isolates of B. xlophilus from China, Portugal, Japan and the US were identical and differed at one to three nucleotides compared to those from Canada. ITS sequences of isolates from China and Portugal were the same; they differed at one or two nucleotides compared to those of Japanese isolates and at four and 23 nucleotides compared to those front the US and Canada, respectively. The phylogenetic analysis indicated that Chinese isolates share a common ancestor with one of the two Japanese clades and that the Canadian isolates form a sister group of the clade comprised of isolates from China, Portugal,Japan, and the US. The relationship between Japanese isolates and those from China was closer than with the American isolates. The Canadian isolates were the basal group of B. xylophilus. This suggests that B. xlophilus originated in North America and that the B. xylphilus that occurs in China could have been first introduced from Japan. Further analysis based on RAPD analysis revealed that the relationship among isolates from Guangdong, Zhejiang, Shandong, Anhui provinces and Nanjing was the closest, which suggests that pine wilt disease in these Chinese locales was probably dispersed from Nanjing, where this disease first occurred in China.

饱食因子及其制备方法和其基因

本发明涉及一种饱食因子及其制备方法和其基因，属于生物医学领域。该饱食因子为从中国两栖类动物大蹼铃蟾(Bombina maxima)皮肤分泌物中分离得到的由28个氨基酸组成的一种单链多肽，分子量3229.57，等电点7.7，多肽氨基酸全序列一级结构为：Asp Met Tyr Glu Ile Lys Gln Tyr Lys The Ala HisGly Arg Pro Pro Ile Cys Ala Pro Gly Glu Gln Cys Pro Ile Trp Val-AMIDATION。制备方法是收集大蹼铃蟾皮肤分泌物，离心去除沉淀、冷冻干燥后，经凝胶过滤、离子交换柱层析、高压液相反相柱层析分离纯化后即得到。编码饱食因子的基因由1041个核苷酸组成，其中编码成熟饱食因子为第835-918位核苷酸。具有抑制进食，增强缓激肽生物效应的作用，作为制备肥胖症治疗药物，心血管系统疾病治疗药物的应用。

Variety of antimicrobial peptides in the Bombina maxima toad and evidence of their rapid diversification

Antimicrobial peptides secreted by the skin of many amphibians play an important role in innate immunity. From two skin cDNA libraries of two individuals of the Chinese red belly toad (Bombina maxima), we identified 56 different antimicrobial peptide cDNA sequences, each of which encodes a precursor peptide that can give rise to two kinds of antimicrobial peptides, maximin and maximin H. Among these cDNA, we found that the mean number of nucleotide substitution per non-synonymous site in both the maximin and maximin H domains significantly exceed the mean number of nucleotide substitution per synonymous site, whereas the same pattern was not observed in other structural regions, such as the signal and propiece peptide regions, suggesting that these antimicrobial peptide genes have been experiencing rapid diversification driven by Darwinian selection. We cloned and sequenced seven genes amplified from skin or liver genomic DNA. These genes have three exons and share the same gene structure, in which both maximin and maximin H are encoded by the third exon. This suggests that alternative splicing and somatic recombination are less likely to play a role in creating the diversity of maximins and maximin Hs. The gene trees based on different domain regions revealed that domain shuffling or gene conversion among these genes might have happened frequently.