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Suk Bong Ha,Da Kun Lee,Kyoung Whan Back 한국응용생명화학회 2002 Journal of Applied Biological Chemistry (J. Appl. Vol.45 No.4
5-Aminolevulinic acid synthase (ALAS) catalyzes the condensation of glycine and succinyl-CoA into 5aminolevulinic acid (ALA) together with the release of CO_2 and CoA. An Escherichia coli expression system was used to generate hexahistidyl-tagged Bradyrhiz
Young-Min Goo,Tae-Won Kim,하선화,Kyoung-Whan Back,Jung-Myung Bae,Yong-Wook Shin,Cheol-Ho Lee,Mi-Jeong Ahn,Shin-Woo Lee 한국식물학회 2009 Journal of Plant Biology Vol.52 No.1
Efforts are being made to identify a parental cultivar suitable for metabolic engineering of potato (Solanum tuberosum L.) that will elevate total carotenoid content or produce a specific type of carotenoid. As an initial step in this search, we performed high performance liquid chromatography analyses and comparisons among gene expression profiles for several cultivars domesticated or bred in South Korea. Here, the dark yellow-fleshed “Golden Valley” contained the highest level of total carotenoids (23.8 μg g−1 dry weight (DW)), which was 1.7- to 3.7-fold higher than those measured in other cultivars. The predominant carotenoids in “Golden Valley” were lutein (40.3% of the total), violaxanthin (29.8%), and β-carotene (8.8%), with only a trace amount of zeaxanthin (0.02%) being detected. Levels of lutein and β-carotene in that cultivar were significantly higher than in the others. Interestingly, relatively high amounts of violaxanthin were accumulated in all cultivars, ranging from 15.9% (1.0 μg g−1 DW in “Jowon”) to 61.7% (8.2 μg g−1 DW in “Dejima”). In accordance with the relatively high content of total carotenoids in “Golden Valley”, remarkably elevated transcripts were also accumulated for most of the genes involved in the carotenoid biosynthetic pathway. In particular, genes encoding enzymes for the first three steps of carotenogenesis—phytoene synthase, phytoene desaturase, and ζ-carotene desaturase—were most actively expressed. A relatively high level of transcript for the carotene hydroxylase (Chy2) gene was detected in all cultivars, including “Jowon”, which had accumulated the lowest amount of total carotenoids. In contrast, almost no transcripts were detected for carotene isomerase (CrtIso) and Chy1 in any of these cultivars. Our preliminary results suggest that “Golden Valley” is an excellent candidate for metabolic engineering that further increases its content of specific carotenoids, e.g., β-carotene and astaxanthin.
Protoporphyrinogen oxidase (Protox) 저해제 저항성 형질전환 벼의 적응성과 Porphyrin 생합성 경로물질과 관련성
윤영범(Young Beom Yun),권오도(Oh Do Kwon),백경환(Kyoung Whan Back),이도진(Do Jin Lee),정하일(Ha Il Jung),국용인(Yong In Kuk) 韓國雜草學會 2011 Weed&Turfgrass Science Vol.31 No.2
Myxococcus Xanthus (MX, PX)와 Arabidopsis thaliana (AP37) 의 protoporphyrinogen oxidase (Protox) 유전자 과다발현 제초제 저항성 형질전환 벼와 비형질전환벼의 생육 및 수량에 관한 적응성 차이와 벼 생육별시기별 ALA 합성능력, tetrapyrrole 중간물질, 활성산소 발생, 지질과산화 작용 및 항산화효소 능력의 연관성이 조사되었다. Protox 과다발현 형질전환 벼 MX와 AP37의 초장은 이앙 후 43, 50, 65일에 비형질전환벼 비형질전환벼에 비해 유의적으로 적었고, 분얼수는 이앙 후 50일과 65일에 MX와 AP37 뿐만 아니라 PX에서도 비형질전환벼에 비해 유의적으로 적었다. 수확기의 간장과 수량은 MX, PX 및 AP37에서 비형질전환벼에 비해 유의적으로 적었다. 형질전환벼 계통 MX, PX 및 AP37 의 수량감소는 MX의 경우 영화수와 천립중에 의해, PX는 등숙율에 의해, AP37은 영화수, 등숙율 및 천립중에 의해서 기인되었다. 한편, 형질전환벼 계통 MX, PX 및 AP37의 수량감소는 또 다른 년찬변이 연구에서도 관찰되었다. 이러한 형질전환벼 계통의 생육 감소는 이앙 후 생육기간 뿐만 아니라 이앙 전 육묘기간 동안에서도 발생하여 결과적으로 수량이 감소되는 것으로 생각된다. Tetrapyrrole 중간물질 Proto IX, Mg-Proto IX 및 Mg-Proto IX monomethyl ester 의 축적량, 활성산소종(H₂O₂ 와 O₂), MDA, 및 항산화효소(superoxide dismutase, caltalase, peroxidase, ascorbate peroxidase, glutathione reductase) 및 엽록소 함량은 Protox 유전자 과다발현 형질전환벼 계통과 비형질전환벼간에 유의적인 차이가 없어 이들 porphyrin 대사 경로 물질과벼 생육 및 수량감소의 연관성은 적은 것으로 사료된다. 그러나 MX, PX, AP37의 ALA 합성능력은 광노출 후 1일과 이앙 후 52일에 비형질전환벼에 비해 유의적으로 감소하여 이 부분에 대한 상세한 연구가 추후에 수행되어야 할 것으로 보이다. The objectives of this study were to investigate fitness difference in growth and rice yield in herbicide-transgenic rice overexpressing Myxococcus xanthus and Arabidopsis thaliana protoporphyrinogen oxidase (Protox) genes and non-transgenic rice. We also aimed to determine whether these fitness differences are related to ALA synthesizing capacity, accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes at different growth stages of rice. Plant height of the transgenic rice overexpressing M. xanthus (MX) and A. thaliana (AP37) Protox genes at 43, 50, and 65 days after transplanting (DAT) was significantly lower than that of WT. Number of tiller of PX as well as MX and AP37 at 50 and 65 DAT was significantly lower than that of WT. At harvest time, culm length and yield of MX, PX and AP37 and rice straw weight of MX and AP37 were significantly low compared with WT. The reduction of yield in MX, PX, and AP37 was caused by spikelets per panicle and 1000 grain weight, ripened grain, spikelets per panicle, 1000 grain weight, and ripened grain, respectively. On the other hand,the reduction of yield in MX, PX, and AP37 was also observed in another yearly variation experiment. The reduction of rice growth in MX, PX, and AP37 was observed in seedling stage as well as growth duration in field. There were no differences in tetrapyrrole intermediate Proto Ⅸ, Mg-Proto Ⅸ and Mg-Proto Ⅸ monomethyl ester, reactive oxygen species (H₂O₂ and O₂-), MDA, antioxidative enzymes (SOD, CAT, POX, APX, and GR) and chlorophyll between transgenic lines and wild type, indicating that accumulated tetrapyrrole intermediate and other parameters were not related to growth reduction in transgenic rice. However, ALA synthesizing capacity in MX, PX, and AP37 at one day after exposure to light and 52 DAT was significantly lower than that of WT. Further study is required to elucidate the mechanisms underlying the growth and yield difference between transgenic and WT lines.
Quality of Rice Lines Transformed with Protox Gene of Bacillus subtilis
Kyung Moon Kim,Yong In Kuk,Wook Kim,Kyoung Whan Back,Ja Ock Guh,Mal Shick Shin 한국육종학회 2006 한국육종학회지 Vol.38 No.1
Characteristics related to grain quality and starch viscosity were investigated in three independent japonica transgenicrice lines expressing a Protox gene from Bacillus subtilis and compared to the control. In this study, we found that there were nosigni