木耳属真菌rDNA特异性扩增片段的RFLP研究

对木耳属8个种25个菌株的ITS和28SrDNA5’端两个区域分别进行了PCR扩增和限制酶切片段长度多态性（RFLP）研究。ITS－RFMP研究结果表明,HaeⅢ可将黑木耳与其它种区分开,MspⅠ可将盾形木耳、角质木耳、琥珀木耳和黑木耳4个种区分开,而供试的HaeⅢ、TaqⅠ、HinfⅠ和MaPⅠ这四种限制酶均不能将皱木耳、大木耳、网脉木耳及毛木耳4个种区分开,表明它们之间的亲缘关系较近;结果还表明,ITS—rDNA拷贝在毛木耳和琥珀木耳种内是异质性的,而在黑木耳种内是同质性的。285rDNA-RFLP研究结果表明,供试的4种限制酶中,仅MspⅠ可将盾形木耳和角质木耳区分开,而不能将其它种区分开,这显示了28SrDNA序列在木耳属不同种间的保守性。     

一株裂解性奶牛乳房炎源大肠杆菌噬菌体的分离鉴定及生物学特性分析

【目的】从新疆石河子地区奶牛粪样中分离裂解性大肠杆菌噬菌体(Escherichia coli phage),对其进行纯化及生物学特性分析。【方法】利用双层平板法从奶牛粪样中分离、纯化噬菌体,将纯化后的噬菌体浓缩液用醋酸双氧铀负染后通过透射电子显微镜观察其形态特征。对该噬菌体进行全基因组测序和遗传进化分析,同时测定噬菌体的宿主谱、最佳感染复数、一步生长曲线、热稳定性及酸碱稳定性。【结果】分离并纯化出一株裂解性噬菌体vB_EcoM_XJ2,噬菌斑圆形不透明,直径0.7 mm–1.2 mm;电镜显示其头部呈正多面体对称,有可伸缩性尾部;核酸类型为双链DNA,基因组大小为75.617 kb,G+C%含量为42.09%;其核酸序列与大肠杆菌噬菌体NJ01和vB_EcoP_SU10相似性高达94%。生物学特性研究显示该噬菌体能裂解多株临床分离的大肠杆菌;能耐受60°C左右高温,在pH 5.0–11.0范围内效价稳定;最佳感染复数为0.1,潜伏期为15 min,暴发期为95 min,裂解量约为10.6 PFU/cell。【结论】vB_EcoM_XJ2是一株在不同温度、不同酸碱性环境中有较强适应能力的裂解性肌尾科大肠杆菌噬菌体。     

海鞘与蓝藻的共生关系

互利共生是指两种不同生物之间所形成的紧密互利关系。文章以海鞘与蓝藻共生关系的典型实例为代表,介绍了蓝藻与海鞘共生关系的发现过程;蓝藻在海鞘间世代传递的机制;两者在共生关系中可能存在的生理功能,如蓝藻共生体通过碳固定、参与氮循环和产生代谢物等方式为海鞘提供营养并参与海鞘的防御活动,海鞘可供给共生体生长所需的一部分氮素营养和提供抗紫外线的保护等。对海鞘与蓝藻共生关系的研究将有助于理解生物进化过程,以及为进一步开发利用海洋生物之间的共生关系提供研究基础。     

利用RAPD技术对木耳属菌株进行分类鉴定的研究

利用ＲＡＰＤ技术对木耳属不同种和种内不同菌株进行了分子鉴定。结果表明,在供试的２６个随机引物中,有１０个引物可扩增出清晰、稳定的ＤＮＡ带型,其中引物Ｓ４和Ｓ６与供试木耳属菌株的基因组ＤＮＡ结合位点少而保守,扩增图谱具有种的特异性,可用于种的快速准确鉴定;其综随机引物的扩增ＤＮＡ多态性较强,可用于种内不同菌株的鉴定;聚类分析表明,在７５％相似水平下,可将供试的木耳属菌株分成四大类,其中有三大类各代表一个形态鉴定的种。研究结果表明了ＲＡＰＤ技术可有效地用于木耳属种或菌株的快速准确鉴定。     

Preliminary research of the RAPD molecular marker-assisted breeding of the edible basidiomycete Stropharia rugoso-annulata

Summary The average RAPD molecular genetic distance was proposed as a criterion in selecting monokaryotic parents for cross breeding and predicting the performance of hybrids of the mushroom Stropharia rugoso-annulata. Three groups of cross pairs or hybrids were recognized based on the average RAPD genetic distance of the monokaryotic parental population. The RAPD-based molecular genetic distance significantly correlated with hybrid mycelial growth rate and mycelial growth heterosis, and their determination coefficients were 0.9237 and 0.8464 respectively. One of the hybrids in group I showed more vigorous mycelial growth in different pH conditions, incubation temperatures, carbon and nitrogen sources, and higher mushroom yield compared with its dikaryotic parent. These results suggested that RAPD-based molecular genetic distance of the monokaryotic parents might be a suitable criterion for selecting monokaryotic parents and predicting the performance of hybrids in mushroom cross breeding.     

Specific inhibition of FAK signaling attenuates subchondral bone deterioration and articular cartilage degeneration during osteoarthritis pathogenesis

Osteoarthritis (OA), a disease of the entire joint, is characterized by abnormal bone remodeling and coalescent degradation of articular cartilage. We have previously found that elevated levels of H-type vessels in subchondral bone correlate with OA and that focal adhesion kinase (FAK) is critical for H-type vessel formation in osteoporosis. However, the potential role of FAK in OA remains unexplored. Here, we demonstrate that the p-FAK level was dramatically elevated in subchondral bone following anterior cruciate ligament transection (ACLT) in rats. Specific inhibition of FAK signaling with Y15 in subchondral bone resulted in the suppression of subchondral bone deterioration and this effect was mediated by H-type vessel-induced ectopic bone formation. Further, articular cartilage degeneration was also alleviated after Y15 treatment. In vitro, the p-FAK level was significantly elevated in mesenchymal stem cells (MSCs) from vehicle-treated ACLT rats as compared to that in MSCs from sham controls and Y15-treated ACLT rats. Elevated p-FAK level in MSCs promoted vascular endothelial growth factor (VEGF) expression, as demonstrated from the high VEGF level in the blood, subchondral bone, and conditioned medium (CM) of MSCs from vehicle-treated ACLT rats. The CM of MSCs from vehicle-treated ACLT rats might promote the angiogenesis of endothelial cells and the catabolic response of chondrocytes through the FAK-growth factor receptor-bound protein 2-mitogen-activated protein kinase-mediated expression of VEGF. The effect of the CM from MSCs of Y15-treated ACLT rats or that treated with a VEGF-neutralizing antibody on vessel formation and the catabolic response was lowered. Thus, the specific inhibition of FAK signaling may be a promising avenue for the prevention or early treatment of OA.     

Retrieving backbone string neighbors provides insights into structural modeling of membrane proteins

Identification of protein structural neighbors to a query is fundamental in structure and function prediction. Here we present BS-align, a systematic method to retrieve backbone string neighbors from primary sequences as templates for protein modeling. The backbone conformation of a protein is represented by the backbone string, as defined in Ramachandran space. The backbone string of a query can be accurately predicted by two innovative technologies: a knowledge-driven sequence alignment and encoding of a backbone string element profile. Then, the predicted backbone string is employed to align against a backbone string database and retrieve a set of backbone string neighbors. The backbone string neighbors were shown to be close to native structures of query proteins. BS-align was successfully employed to predict models of 10 membrane proteins with lengths ranging between 229 and 595 residues, and whose high-resolution structural determinations were difficult to elucidate both by experiment and prediction. The obtained TM-scores and root mean square deviations of the models confirmed that the models based on the backbone string neighbors retrieved by the BS-align were very close to the native membrane structures although the query and the neighbor shared a very low sequence identity. The backbone string system represents a new road for the prediction of protein structure from sequence, and suggests that the similarity of the backbone string would be more informative than describing a protein as belonging to a fold.     

RAPD molecular differentiation of the cultivated strains of the jelly mushrooms, <Emphasis Type="Italic">Auricularia auricula</Emphasis> and <Emphasis Type="Italic">A. polytricha</Emphasis>

Auricularia mushrooms are the fourth most important cultivated mushrooms in the world, with a unique jelly taste and horizontal-septated basidium which are significantly different from other cultivated mushrooms. Differentiation of commercial cultivated strains is difficult to conduct, due to the lack of useful distinguishable characters. In this study, we used the RAPD technique to differentiate 11 commercial strains of A. auricula and five commercial strains of A. polytricha and one white-fruitbody mutant strain, and to characterize their genetic diversity. Results showed that all the strains tested could be differentiated by pooled RAPD data, and even one individual primer (S10) could also discriminate all tested strains. RAPD analysis could differentiate strains having identical rDNA RFLP and supports the classification of the white-fruitbody mutant to the species of A. polytricha. Genetic similarity analysis and grouping derived from RAPD markers reveals a high level of genetic diversity of commercial strains of Auricularia auricula and A. polytricha. Therefore, the RAPD technique can provide a powerful tool to discriminate the commercialAuricularia strains and offer the molecular information useful for breeding systems.