Expression of Aspergillus niger N5-5 in E. coli and purification and identification of products

Due to the feature of high hydrolysis, tannase is widely used in food, beverage, brewing and other fields. However, high cost in producing natural tannase makes it difficult to apply tannase to industry in a large-scale. Microbial expression systems can be used for preparing numerous amount of enzyme at low cost, so in this paper Aspergillus niger N5-5 was expressed using E. coli system. Specific primers were designed based on the Aspergillus niger N5-5 sequence N3 (GenBank, No.: KP677552), and tannase gene tan was promoted to carry 6 His tag and enzyme cutting site which contains NdeI/HindIII using PCR amplification. Then, tannase gene tan was connected to expression vector by NdeI/HindIII enzyme cutting. In this way, recombinant expression vector tan-pET43.1a was formed. Then, the expression vector pET43.1a by NdeI/HindIII enzyme cutting was transformed into E. coli BL21 (DE3) to induce expression of Aspergillus niger N5-5. When the induced fungi were disrupted by the ultrasonic wave, the crude enzyme was extracted and purified by using the IMAC, and then the activity of the crude enzyme and pure enzyme was determined. According to the results of determination of the tannase activity, the tannase activity of the crude enzyme was greatly improved after the crude enzyme was purified, and the specific activity of the pure enzyme was about 8 times of that of the crude enzyme. The results of SDS-PAGE of the pure enzyme showed that the molecular mass of the pure enzyme was about 65 kDa/64–65 kDa, which was consistent with the expected result (64.2 kDa), It can be concluded that the crude enzyme solution was purified successfully. The results of pure enzyme’s protein identification by Western Blotting showed that clear protein bands pro-3 were observed. Molecular mass of clear protein bands pro-3 was about 65 kDa, which was in line with the expected results (64.2 kDa). It can be seen that the aforementioned expression protein could be specifically combined with His tag. It proved expression protein to be a recombinant fusion protein with 6 His tag.     

Review of the New Caledonian species of Acritoptila Wells, 1982 (Trichoptera,Insecta), with descriptions of 3 new species

We review the New Caledonian representatives of the Australasian endemic hydroptiline genus Acritoptila, based on examination of a considerable collection of material in the Swedish Museum of Natural History and of types of previously established species. A key for identification of males is given and includes 3 species newly described here: A. parallela sp. n., A. forficata sp. n. and A. macrospina sp. n. For all New Caledonian species, male genitalia are illustrated, and for 5 associated females, distinctive features are illustrated and described.     

Succession in zooplankton and hydrophytes of a seasonal water on the west coast of South Africa

Paramesochra mielkei sp.n. is described and figured from the interstices of subtidal sandy sediments off the SW Dutch coast. Kunz' (1981) phylogenetic scheme of the Paramesochridae Lang, 1948 is re-examined and it is suggested that the family comprises two phyletic lines which originated early in paramesochrid evolution. Translation into Linnean hierarchies implies the establishment of two new sub-families. Within the primitive Diarthrodellinae subfam. n., Tisbisoma Bozic, 1964 is ancestral to Diarthrodella Klie, 1949 s.l. and Rossopsyllus Soyer, 1975. Remanea Klie, 1929 is transferred to the Paramesochrinae subfam. n. which comprises the genera of both the Scottopsyllus- and the Paramesochra-group. The aberrant genus Caligopsyllus Kunz, 1975, standing close to Apodopsyllus, is removed from the Paramesochra-group. P. brevifurca Galhano, 1970 is splitted into two subspecies and replaced in the genus Paramesochra. An attempt is made to assess the phyletic interrelationships of the Paramesochra-species and the resulting cladogram splits the genus into four species-groups. P. mielkei sp.n. is referred to the dubia-group and seems to be closely related to P. borealis Geddes, 1981. Finally, an amended diagnosis and a revised key to the species of the genus Paramesochra are presented.     

中药马蓝叶及其混乱品的比较鉴别

马蓝叶是加工传统中药青黛的主要原料。为了确保药材质量,作者对中药马蓝叶及其混乱品种球花马蓝、疏花马蓝、广西马蓝的原植物、药材性状、叶组织显微特征和薄层层析鉴别等进行了研究。指出其混乱品种虽与马蓝叶是同属植物,但不含马蓝叶的有效成分靛玉红,应仔细鉴别,不宜混用。     

酵母鉴定程序在酵母分类鉴定中的应用

本文介绍了Barnett等1985年编制并由剑桥大学发行的计算机软件《酵母鉴定程序》,以及如何使用该程序在IBM PC DOS操作系统上进行酵母菌的分类鉴定。我们使用该程序对从云南鸡足山和紫金山两地森林土壤中分离到的82株酵母菌进行了分类鉴定,其中:鉴定到种的有47株,占总株数的57.3％;鉴定到属但未能直接定到种的有21株,占总株数的25.6％;暂未定名的有14株,占总株数的17.1％。     

腐霉属分类性状评价及其中国的种

本文对腐霉属Pythium Pringsheim的研究历史作了简单的回顾,对该属的分类性状和系统进行了论述和评价,最后对中国已发现的55种腐霉,以检索表的方式进行了分类、检索。     

The systematics and ecology of snakeheads (Pisces: Channidae) in Peninsular Malaysia and Singapore

Seven species of snakeheads (Channidae) are known from Peninsular Malaysia and Singapore, viz. Channa bankanensis, C. gachua, C. lucius, C. marulioides, C. melasoma, C. micropeltes and C. striata. Up-to-date distribution maps of each species are presented, including new records. Their systematics is reviewed and partially revised. The taxonomic status of C. marulioides and C. melanoptera is clarified. Specimens from Peninsular Malaysia identified as C. melanoptera sensu Weber & de Beaufort, 1922, proved to be the adult form of C. marulioides s.str. The real C. melanoptera appears to be restricted to Borneo and possibly Sumatra. The life history of a blackwater species C. bankanensis is also documented, with regards to the morphological and colour-pattern changes associated with growth. An updated key to the seven species based on morphometric measurements and meristic counts is presented.     

中国瑟姬小蜂及二新种的描述（膜翅目：姬小蜂科）

瑟姬小蜂属Cirrospilus昆虫是鳞翅目,双翅目,特别是细蛾,潜叶蛾,潜叶蝇类及其他荫蔽性小幼虫或蛹的重要寄生蜂,有些也能重寄生于绒茧蜂和蜘蛛卵等中,本文除二新种描述外,还收入瑟姬小蜂六种。一并列于检索表中。     

我国大麦和性花叶病毒（BaMMV）研究初报

本文根据F(ab′)2－ELISA和ISEM实验,结合病毒粒子长度测定,报道了江苏省如东县农科所大麦黄花病毒源（吕种盐辐矮早3）中除存在大麦花花叶病毒（BaYMV）外,还复合感染大麦和性花叶病毒（BaMMV）,这是我国BaMMV的第一次报道。