生物安全学报
生物安全學報
생물안전학보
Journal of Biosafety
2015年
2期
18-31
,共14页
申建茹%Ying YAN%武强%李建伟%张桂芬%万方浩
申建茹%Ying YAN%武彊%李建偉%張桂芬%萬方浩
신건여%Ying YAN%무강%리건위%장계분%만방호
遗传修饰昆虫%转化标记%眼睛颜色标记%抗药性标记%荧光蛋白标记
遺傳脩飾昆蟲%轉化標記%眼睛顏色標記%抗藥性標記%熒光蛋白標記
유전수식곤충%전화표기%안정안색표기%항약성표기%형광단백표기
genetically modified insect%transformation marker%eye color gene%drug resistance gene%fluorescent protein gene
遗传转化标记是将遗传修饰昆虫从野生型种群中分辨出来的根据,遗传转化昆虫的鉴定、转化品系的维持及其遗传稳定性的监测都依赖于可靠的标记系统,发展易于应用和监测的转化标记能够极大地促进害虫遗传防治的相关研究。用于遗传修饰昆虫的转化标记主要有昆虫眼睛颜色标记基因、抗药性标记基因和荧光蛋白标记基因等。非果蝇类昆虫首个遗传转化品系的鉴定是通过眼睛颜色突变而实现,但大多数昆虫物种没有可用的突变体或缺少相应基因的信息,从而限制了眼睛颜色标记的应用。抗药性基因标记虽然能够通过对转化昆虫进行集体选择而大幅度提高筛选转化体的效率,但由于其鉴定的准确性不高且存在安全性问题,未得到广泛应用。荧光蛋白标记基因的发展则显著拓宽了能够转化的昆虫种类。从水母分离的绿色荧光蛋白( GFP )经突变方法获得了多种不同荧光性质的突变体,经人为修饰后与适宜的强启动子构成转化标记载体,能够有效鉴定更多昆虫物种的遗传转化个体,其中应用较多的是增强型绿色荧光蛋白( EGFP )。此外,从珊瑚属海葵中分离得到的红色DsRed标记基因提供了多样化的红色荧光蛋白选择,在某些生物中DsRed与GFP联合应用的表现明显优于GFP突变体,所以其应用前景也非常广泛。本文着重从眼睛颜色、抗药性和荧光蛋白等3个方面阐述了标记基因的发展历史与现状,并对其今后的发展方向进行了展望。
遺傳轉化標記是將遺傳脩飾昆蟲從野生型種群中分辨齣來的根據,遺傳轉化昆蟲的鑒定、轉化品繫的維持及其遺傳穩定性的鑑測都依賴于可靠的標記繫統,髮展易于應用和鑑測的轉化標記能夠極大地促進害蟲遺傳防治的相關研究。用于遺傳脩飾昆蟲的轉化標記主要有昆蟲眼睛顏色標記基因、抗藥性標記基因和熒光蛋白標記基因等。非果蠅類昆蟲首箇遺傳轉化品繫的鑒定是通過眼睛顏色突變而實現,但大多數昆蟲物種沒有可用的突變體或缺少相應基因的信息,從而限製瞭眼睛顏色標記的應用。抗藥性基因標記雖然能夠通過對轉化昆蟲進行集體選擇而大幅度提高篩選轉化體的效率,但由于其鑒定的準確性不高且存在安全性問題,未得到廣汎應用。熒光蛋白標記基因的髮展則顯著拓寬瞭能夠轉化的昆蟲種類。從水母分離的綠色熒光蛋白( GFP )經突變方法穫得瞭多種不同熒光性質的突變體,經人為脩飾後與適宜的彊啟動子構成轉化標記載體,能夠有效鑒定更多昆蟲物種的遺傳轉化箇體,其中應用較多的是增彊型綠色熒光蛋白( EGFP )。此外,從珊瑚屬海葵中分離得到的紅色DsRed標記基因提供瞭多樣化的紅色熒光蛋白選擇,在某些生物中DsRed與GFP聯閤應用的錶現明顯優于GFP突變體,所以其應用前景也非常廣汎。本文著重從眼睛顏色、抗藥性和熒光蛋白等3箇方麵闡述瞭標記基因的髮展歷史與現狀,併對其今後的髮展方嚮進行瞭展望。
유전전화표기시장유전수식곤충종야생형충군중분변출래적근거,유전전화곤충적감정、전화품계적유지급기유전은정성적감측도의뢰우가고적표기계통,발전역우응용화감측적전화표기능구겁대지촉진해충유전방치적상관연구。용우유전수식곤충적전화표기주요유곤충안정안색표기기인、항약성표기기인화형광단백표기기인등。비과승류곤충수개유전전화품계적감정시통과안정안색돌변이실현,단대다수곤충물충몰유가용적돌변체혹결소상응기인적신식,종이한제료안정안색표기적응용。항약성기인표기수연능구통과대전화곤충진행집체선택이대폭도제고사선전화체적효솔,단유우기감정적준학성불고차존재안전성문제,미득도엄범응용。형광단백표기기인적발전칙현저탁관료능구전화적곤충충류。종수모분리적록색형광단백( GFP )경돌변방법획득료다충불동형광성질적돌변체,경인위수식후여괄의적강계동자구성전화표기재체,능구유효감정경다곤충물충적유전전화개체,기중응용교다적시증강형록색형광단백( EGFP )。차외,종산호속해규중분리득도적홍색DsRed표기기인제공료다양화적홍색형광단백선택,재모사생물중DsRed여GFP연합응용적표현명현우우GFP돌변체,소이기응용전경야비상엄범。본문착중종안정안색、항약성화형광단백등3개방면천술료표기기인적발전역사여현상,병대기금후적발전방향진행료전망。
Transformation markers offer a tool to distinguish the genetically modified insects from wild types. Both the identification of transformants and the maintenance of transformed lines depend on reliable transformation makers. In addition, the evaluation of the ge-netic stability of released genetically modified insects needs strong and stable markers. Thus the development of broadly applicable, easily detectable and reliable transformation markers will facilitate the study of genetic pest management. In general, eye color genes, drug resistance genes and fluorescent protein genes can be used as markers in genetically modified insects. The first efficient identifica-tion of a non-drosophilid insect transformation line was based on the rescue of eye color mutant phenotypes. However, for most insect species, the application of eye color markers is limited because of the lack of suitable recipient mutant strains and less information on related genes. Markers based on drug resistance genes can improve the screening efficiency of transformants, but the selection for drug resistance is problematic and prone to have false positives or negatives with potential biosecurity problems. Fluorescent protein gene markers significantly facilitate the development of stable insect transformation lines. The green fluorescent protein (GFP, isolated from the jellyfish Aequorea victoria) and its variants with various fluorescent characteristics can be combined with suitable, strong promoters to serve as transformation markers for a wide range of insect species and guarantee the reliable screening of the transformants. In this category, the enhanced green fluorescent protein (EGFP) was mostly used. Besides, the red fluorescent protein (DsRed), isolated from the mushroom coral, Discosoma striata, provides a selection of red fluorescent proteins with better performance than GFP mutants. This paper reviews the history and status of transformation markers including eye color genes, drug resistance genes and the fluorescent protein genes. The potential roles of transformation markers in genetic pest management are also discussed.