Spider silks hold great potential as biomaterials with extraordinary properties. Here we report cloning and characterization of the major ampullate silk protein gene from the spider Araneus ventricosus. A cDNA coding for the partial major ampullate silk protein (AvMaSp) was cloned from A. ventricosus. Analysis of the cDNA sequence shows that AvMaSp consists of 240 amino acids of a repetitive region and 99 amino acids of a C-terminal non-repetitive domain. The peptide motifs found in spider major ampullate silk proteins, (A)n, (GA)n, and (GGX)n, were conserved in the repetitive region of AvMaSp. Phylogenetic analysis further confirmed that AvMaSp belongs to the spider major ampullate spidroin proteins. The AvMaSp-R cDNA, which contains sequences encoding for 240 amino acids of a repetitive domain, was expressed as a 22 kDa polypeptide of soluble form in baculovirus-infected insect cells. Recombinant AvMaSp-R was degraded abruptly by trypsin. However, AvMaSp-R was stable at 100 °C for at least 30 min. Additionally, the AvMaSp-R was stable at various pH values from 2 to 12 for at least 1 h. Taken together, our findings provide the molecular structure and biochemical property for A. ventricosus major ampullate silk protein as a biomaterial.

Müllerian inhibiting substance (MIS) is a protein that encoded by MIS gene. It has also been called Müllerian inhibiting factor (MIF) and anti-Müllerian hormone (AMH). Mis expression occurs in ovarian granulose cells of females postpartum, and serves as a molecular biomarker for relative size of the ovarian reserve. In humans, the number of cells in the follicular reserve can be used to evaluate the reproductive function and fertility of female. Pagrus major is typical cultured fish in Korea but there is no clear evidence for their gene identification. However, in many teleost, MIS genes were demonstrated already. Present study aimed to identify the Mis gene in Pagrus major and seasonal difference of its expression. Using conserved sequence of the other known teleost Mis genes, we make conserved primers. Pagrus major’s ovary samples were obtained from the sea rim farm (Geoje, Korea) and kept in RNAlater®Solution or fixed in 4% paraformaldehyde containing 0.16% picric acid. RNA was isolated from kept sample and cDNA was synthesized. The PCR products were performed ligation with TOPO vector and transformation in TOP10 cell and sequenced the Mis mRNA fragment. MIS was localized in the follicle cells. Its mRNA levels were higher in summer than spring or fall. Based on them, it is suggested that MIS can be used to estimate the fertility of this fish.

Fibroin silk proteins from spider or silkworm are attractive biomaterials that are of particular biotechnological interest for industrial and medical purposes because of their unique physical and mechanical properties. In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. Spider silks have great potential as biomaterials with extraordinary properties. Here, we report the cloning and characterization of the major ampullate silk protein gene from the spider Araneus ventricosus. A cDNA encoding the partial major ampullate silk protein (AvMaSp) was cloned from A. ventricosus. An analysis of the cDNA sequence shows that AvMaSp consists of a 240 amino acid repetitive region and a 99 amino acid C-terminal non-repetitive domain. The peptide motifs that were found in the spider major ampullate silk proteins, (A)n, (GA)n, and (GGX)n, were conserved in the repetitive region of AvMaSp. Phylogenetic analysis further confirmed that AvMaSp belongs to the spider major ampullate spidroin family of proteins. Recombinant AvMaSp-R was degraded abruptly by trypsin. However, AvMaSp-R was stable at 100 °C for at least 30 min. Additionally, the AvMaSp-R was stable at pH values from 2 to 12 for at least 1 h. Taken together, our findings describe the molecular structure and biochemical properties of the A. ventricosus major ampullate silk protein and demonstrate its potential as a biomaterial.

In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. A cDNA coding for the C-terminus of spider silk protein (AvMaSp) was cloned from the spider Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvMaSp consists of 165 amino acids of are petitive region and 99 amino acids of a C-terminalnon-repetitive region. The peptide motifs found in spider silk proteins, GGX and An, were conserved in the repetitive region of AvDrag. The AvMaSp cDNA was expressed as a 28kDa polypeptide in baculovirus-infected insect cells. To produce transgenic rice plant with high contents of glycine and alanine, the prolamin promoter-driven AvMaSp was introduced into rice plant via Agrobacteriumtumefaciens-mediated gene transformation. Because of seeds pecific prolamin promoter, expression of AvMaSp protein has been achieved inriceseed. The introduction and copy number of the AvMaSp gene in transgenic rice plants were determined by PCR and Southern blot analysis. AvMaSp expression in transgenic rice seeds was examined by Northern blot and Western blot analysis. Immuno fluorescence staining with the AvMaSpantiserum revealed that the recombinant AvMaSp proteins were localized in transgenic rice seeds. Furthermore, the amino acid content analysis showed that transgenic rice seeds were greatly increased in glycine and alanine as compared to controls. The present study is the first to show the expression of spider silk protein in rice seed.

A gene flow has been tested from transgenic herbicide-resistant Chinese cabbage to major crops in Cruciferae. Hybridizations were made between transgenic Basta-resistant Chinese cabbage and non-transgenic Chinese cabbage, B. napus, B. juncea and Brassicoraphanus(B. campestris × Raphanus sativus, 2n=4x=38) with honey-bee mediated fertilization. Progeny populations were selected by Basta herbicide. The F1 hybrids with pat gene were again self-pollinated or cross-pollinated with nontransgenic Chinese cabbage and rape naturally or artificially. Pod setting ratio were compared as a tentative parameter for potential gene flow. Key agronomic characters were compared among the F1 hybrid populations obtained. In the days to flowering after cold treatment, the F1 hybrids of Brassicoraphanus x B. campestris and B. napus x B. campestris(GM) were the shortest (75 days), and the GM Chinese cabbage and the F1 hybrid of B. juncea x B. campestris(GM) were 87 and 93 days, respectively. In the stem length, the F1 hybrid of Brassicoraphanus x B. campestris was the longest(175 cm), and followed by the F1 hybrid of B. napus x B. campestris(GM), the F1 hybrid of B. juncea x B. campestris(GM) in order. The naturally occurring pod setting ratios were fluctuated seriously among individual and segregation lines. Natural pod setting of the transgenic Chinese cabbage was 10 to 19 %. The F1 hybrid of B. napus x B. campestris(GM) showed the ratio of 4-11%. However, the F1 hybrids of B. juncea x B. campestris and Brassicoraphanus x B. campestris(GM) did not produced seeds. The F1 hybrids with pat gene were hybridized with nontransgenic Chinese cabbage and rape by artificial bud pollination to test potential gene flow. All cross combinations except Brassicoraphanus produced seeds and the pod setting ratios ranged from 35 to 100 percentages. Thus, the potentials of the gene flow from Chinese cabbage with pat gene to the order specise in the same gene interspecies or intergeneric species and from the F1 hybrids with pat gene to nontransgenic Chinese cabbage and rape were detected. Herbicide resistant test to the acquired seeds by natural and artificial pollinations including honey-bee mediated fertilization using the same materials is going on.

Two important loci, designated Rhg1 and Rhg4, are responsible for most resistance to soybean cyst nematode (SCN). The objective of this study was to survey single nucleotide polymorphism (SNP) and to map the SCN-resistant gene, Rhg4, on soybean genetic ma