Construction and Expression of Human Survivin and Preparation of Its Polyclonal Antibody

Survivin, a novel member of inhibitor of apoptosis（IAP） protein family, is aberrantly expressed in cancer but undetectable in normal, differentiated adult tissues. The cancer-specific expression of survivin, coupled with its importance in inhibiting cell death and in regulating cell division makes it a useful diagnostic marker of cancer and a potential target for cancer treatment. Survivin cDNA amplified from the total RNA of 293 cells through RT-PCR was cloned into prokaryotic expression vector pRSET-B. The recombinant plasmid pRSET-B-Surv was expressed in E.coli BL21, and the relative molecule mass（Mr） of expressed fusion protein was approximately 21000. The recombinant protein was purified through Ni^2＋ affinity chromatography column and characterized by SDS-PAGE and Western blot. The purified recombinant protein was then injected into rabbits, and antisurvivin polyclonal antibody with a high titer was obtained.     

The join of two minimal clones and the meet of two maximal clones

We construct two minimal clones on any finite set such that the join of the two clones contains all operations. Dually, we exhibit two maximal clones on any finite set with at least three elements such that the intersection of the two clones is the trivial clone containing projections only. Received October 20, 1998; accepted in final form December 16, 1998.     

A statistical model for global optimization by means of select and clone

A statistical model tor giobai optimization is constructed generalizing some properties ofthe Wiener process to the multidimensional case. A new approach, which is similar to the Branch and Bound approach, is proposed to the construction of algorithms based on statistical models. A two dimensional version of the algorithm is implemented, and test results are presented     

Evaluation of thermal behavior of latex membranes from genetically improved rubber tree (Hevea brasiliensis)

Thermal stability, thermal decomposition process, residual mass, temperature of glass transition (T_g) and temperature dependence of storage modulus (E’), were determined for latex membranes prepared from six clones of Hevea brasiliensis: IAC 331, IAC 332, IAC 333 and IAC 334 grown at experimental plantations of Instituto Agronômico de Campinas (IAC) in Votuporanga, São Paulo State, Brazil. Latex membranes from GT1 and RRIM 600 Asian matrix clones were used as references. The thermal behavior of latex membranes from genetically improved rubber trees was characterized using thermogravimetry/derivative thermogravimetry (TG/DTG), differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA). The thermal behavior of latex from clones studied in the present work showed similar features of the clones previously reported (IAC 40, IAC 300, IAC 301, IAC 328, IAC 329 and IAC 330), with mass loss in four consecutive steps, except IAC 333, which showed an additional mass loss step.     

Method for clone checking

A new method for simple and fast clone checking is described. We combined the Pyrosequencing technology with a preprogrammed nucleotide dispensation strategy for fast analysis of DNA constructs. To test this method, the N-terminus region of plasmids constructed for the production of recombinant apyrase was analyzed. Of the ten plasmids tested, seven constructs were correct, two constructs showed one base deletion, and one construct showed deletion of a 195 bp fragment. The preprogrammed nucleotide dispensation strategy allowed the identification of the sequence downstream of the deletions. Thus, this method determines both the location and nature of possible artifacts.     

On clones, transformation monoids, and finite Boolean algebras

We prove that, for a finite Boolean algebra, there exist only finitely many clones which consist of polynomial functions of the algebra and contain the monoid of its unary polynomial functions. Received January 7, 2000; accepted in final form December 13, 2000.     

SPAP2基因的克隆、融合表达及分离纯化

本文构建的表达载体pGex-2T-SPAP2CT在大肠杆菌中表达出可融性蛋白, 其分子量为46 000, 经纯化后得到产率为10%、 纯度大于90%的GST-SPAP2CT蛋白.     

A Generalized Version of the Baker–Pixley Theorem

A famous theorem by Baker and Pixley states that the term functions of a finite algebra with a (d + 1)-ary near-unanimity term are precisely the functions under which all subalgebras of the algebra's dth power are closed. In this paper, we generalize the theorem to a completely abstract level. Indeed, we obtain a version of the theorem that is stated in purely category-theoretic terms, making it applicable in any concrete or abstract category. To motivate this rather abstract result, we also discuss some of its concrete applications.     

Binary operations suffice to test collapsing of monoidal intervals

It will be shown that the stabilizer clone of a transformation monoid is either trivial, i.e., it is generated by the monoid itself, or it contains an essentially binary function. Received September 16, 1996; accepted in final form May 21, 1997.     

cDNA cloning,expression and bioinformatical analysis of Tssk genes in tree shrews

Tree shrews are more closely related to primate animals than rodents in many aspects.In addition, they also possess several advantageous characteristics including small body size, high brain-to-body mass ratio, low cost of feeding and maintenance, short reproductive cycle and life span, which make them promising novel laboratory animals to replace more precious larger primate animals. Testis-specific serine/threonine kinase (Tssk) plays important roles in spermatogenesis and/or the regulation of sperm function. However, studies on Tssk in tree shrews have not been reported yet. In the present study, the full-length sequences of five members of the Tssk family in tree shrews were cloned and their CDS region sequences were analyzed by basic bioinformatics. The phylogenetic tree and prokaryotic protein expression system of Tssk gene of tree shrews were constructed. The mRNA expressions of Tssk genes in 11 tissues/organs from tree shrews were studied. The results showed that: 1. the length of the CDS region of tree shrew Tssk gene for Tssk1B, Tssk2, Tssk3 (variant X1 / X2), Tssk4 (variant X1 / X2) and Tssk6 is 1080bp, 1077bp, 867 / 807bp, 1014 / 984bp, 822bp, respectively, encoding 359, 358, 288/268, 337/327 and 273 amino acids, respectively; the cloned sequences of Tssk genes have been submitted to GenBank with the following accession numbers: KX091161(Tssk1B), KX091162(Tssk2), KX091163(Tssk3 variant X1)/KX091164(Tssk3 variant X2), KX091165(Tssk4variant X1)/KX091166(Tssk4variant X2), KX091160(Tssk6). 2. All tree shrew Tssk proteins distribute in cytoplasm, indicating that they are hydrophilic and non-secretory proteins, with multiple phosphorylation sites of serine and/or threonine. In addition, they are all mixed proteins with similar tertiary structures sharing a highly conserved functional domain of S_TKc (Serine/Threonine protein kinases, catalytic domain). 3.The molecular phylogenetic tree of five Tssk genes in tree shrews indicates that they are neither rodent nor primate animal, but are closely related to primate animals. 4. Five members of the Tssk recombinant proteins in tree shrews were successfully obtained using the constructed prokaryotic protein expression system. 5. Five Tssk genes are specifically expressed in the testis and/or sperm of tree shrews. Additionally, small amount of Tssk1B was expressed in several tissues other than testis and sperm. Limited mRNA levels of Tssk2 and Tssk4 were expressed in the brain, while mRNA of Tssk3 or Tssk6 could only be detected in the testis and sperm.This study will provide fundamental data on reproductive biology of tree shrews, which paves a way for further studying Tssk’s biological function in this novel model animal.