Bt crystal proteins, encoded by cry genes, are a group of insecticidal proteins unique in the Gram-positive and spore-forming bacterium, Bacillus thuringiensis. These cry genes are widely applied as one of the most successful candidates for constructing transgenic plants resistant to pest insects. In our previous report, we found Cry1-5 had high insecticidal activity against Spodoptera larvae although its amino acid sequences showed high similarity (97.9%) to those of Cry1Ab which had low activity. In comparison with Cry1Ac, Cry1-5 had 12 different residues in domain Ⅰ and domain Ⅱ, and we focused on domain Ⅰand domain Ⅱ regions and designed 10 mutagenic primers to change 12 residues. Through multi site-directed mutagenesis, we mutated the modified cry1Ac gene by plant codon usage in pOBⅠ-Mod-cry1Ac based on cry1-5 and constructed 63 various mutant cry genes. In the further study, we will express those mutant proteins as a fusion form with polyhedrin using baculovirus expression system and subsequently do bioassay to Spodoptera larvae.

A strain of Bacillus thuringiensis, named Bt 1-3, was isolated from Korean soil sample and it showed high insecticidal activity against Plutella xylostella. Bt 1-3 was deterimined to belong to ssp. aizawai (H7) by an H antiserum agglutination test and produced bipyramidal-shaped crystal proteins. PCR analysis with specific cry gene primers showed that Bt 1-3 contained cry1Aa, cry1Ab, cry1C, cry1D and cry2Ab genes. In addition, this isolate showed high uptake rate of foreign plasmid by electroporation. Based on these characteristics of Bt 1-3, we tried to construct a spore-free Bt 1-3 mutant by knock-out sigG gene, which is known as a key transcription factor during sporulation. First, we constructed a basal vector, named pDST, consisting of erythromycin resistant gene (EmR), partial polyhedrin gene and temperature sensitive origin of replication gene (Orits). Subsequently, according to the chromosomal DNA sequence of Bt subsp. konkukian 97-27, we amplifed upstream and downstream regions of Bt 1-3 sigG, and cloned into pDST (pDST-G). So far, several EmR colonies were obtained by electroporating into the wildtype Bt 1-3 and crossover by homologous recombination is going on.

To develop an improved baculovirus insecticide with additional advantages, a novel recombinant baculovirus, AcB5B-AaIT was constructed. B. thuringiensis crystal protein gene (cry1-5) and insect-specific neurotoxin gene (AaIT) were introduced into Autographa californica nucleopolyhedrovirus genome by fusion of polyhedrin-cry1-5-polyhedrin under the control of polyhedrin (polh) gene promoter, and AaIT under the control of early promoter of ORF3004 from Cotesia plutellae bracovirus, respectively. About 150 kDa of Polyhedrin-Cry1-5-Polyhedrin fusion protein expressed by AcB5B-AaIT was occluded into the polyhedra produced by the recombinant virus, and activated as about 65 kDa of crystal protein when treated with gut-juice of Bombyx mori. The AcB5B-AaIT showed about 50% reduced LT50 value compared to that of the recombinant virus, Ap1Ac, expressing Cry1Ac against Plutella xylostella larvae. In addition, Spodoptera exigua larvae fed the recombinant polyhedra of AcB5B-AaIT showed about 4 fold higher refusing diet effect compared S. exigua larvae fed the recombinant polyhedra of the recombinant virus, Ap1C, expressing Cry1C. AcB5B-AaIT could be transferred to wild-type baculovirus along with serial passage by the homologous recombination between two polyhedrin genes contained in polh-cry1-5-polh fusion protein gene. These results suggested that the novel recombinant baculovirus, AcB5B-AaIT, could be applied as advanced viral insecticide.

Antimicrobial stability of grape seed extract (ActiVinTM), pine bark extract (Pycnogenol®), and oleoresin rosemary (Herbalox®) on the growth of Aeromonas hydrophila was investigated in cooked ground beef. When compared to the control, the populations of A. hydrophila were most effectively reduced by 4.06 log CFU/g for 1% Pycnogenol® added after cooking at 10 days of refrigerated storage, followed by 3.06 log CFU/g for 1% Pycnogenol® added before cooking and 1.36 log CFU/g for ActiVinTM. Bacteriostatic and bactericidal activities were observed for Pycnogenol® added before and after cooking, respectively. Pycnogenol® consists of heat-labile and heat-stable compounds. ActiVinTM and Pycnogenol® could be considered for use as multifunctional preservatives in meat and meat products.

The removal of oxygen during sintering by carbothermic reduction was studied for steel compacts Fe-Cr-Mo-C and Fe-Mo-C prepared from prealloyed powders. The compacts were prepared by pressing at 600 and 1000 MPa and sintering at 1100 and 1300°C in vacuum. It showed that for the Cr-Mo steel, deoxidation strongly depends on the sintering temperature, in contrast to the plain Mo steel; at 1300°C very low oxygen levels were measured with the standard density compact while at high density still significant oxygen is contained. This indicates inhibition of final deoxidation by pore closure, but apparently without adverse effect on the mechanical properties.

Cloning by nuclear transfer in mammals using somatic cells has enormous potential applications. However, somatic cloning has been inefficient in all species in which NT is successful. High abortion and fetal death rates have been observed. These developmental defects have been attributed to incomplete nuclear reprogramming by the somatic cloning process. In this review, we will discuss studies conducted in our labs to understand the nuclear reprogramming process.

LAMOST is a special reflecting Schmidt telescope. Both the refleting Schmidt plate MA and the spherical primary mitrror MB are segmened mirrors. These two cofocus but not co-phase. The diffraction of the optical system is decided by the shape overlapping of MA and MB. This paper describes the diffraction caculating results with different declination and different field angle. The diffraction influence to the image quality is acceptable in the error buget of optical system. It also proves that the size seletion of the sub-mirror is reasonable.