中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2012年
11期
961-966
,共6页
梁宇%靳玉琴%张靖%陈实%李启明
樑宇%靳玉琴%張靖%陳實%李啟明
량우%근옥금%장정%진실%리계명
拟多形假丝酵母菌%营养缺陷型%ura3基因%leu2基因
擬多形假絲酵母菌%營養缺陷型%ura3基因%leu2基因
의다형가사효모균%영양결함형%ura3기인%leu2기인
Candida parapolymorpha%Auxotrophic%ura3 gene%Leu2 gene
目的 以拟多形假丝酵母菌(ATCC26012)为材料,构建遗传稳定性良好的尿嘧啶和亮氨酸双营养缺陷型工程化菌株.方法 利用物化及基因工程等方法对拟多形假丝酵母菌株的染色体进行改造,将ura3基因和leu2基因进行定向诱变,以获得尿嘧啶和亮氨酸双营养缺陷型突变菌株,并对其进行生物学鉴定,包括菌株的遗传稳定性、基因序列及生理生化特征分析等.结果 筛选得到尿嘧啶和亮氨酸双营养缺陷型突变菌株,鉴定结果显示该菌株遗传稳定性良好,标志基因均发生定向改变,生理生化结果显示该菌株能利用多种碳源和氮源,且生长状况良好.结论 拟多形假丝酵母的双营养缺陷型工程化改造为进一步研究该菌株的遗传学奠定了基础,为多价蛋白组成类病毒颗粒的外源表达提供了工程化宿主.进而,为复杂疫苗抗原的重组表达和功能研究提供了技术手段.
目的 以擬多形假絲酵母菌(ATCC26012)為材料,構建遺傳穩定性良好的尿嘧啶和亮氨痠雙營養缺陷型工程化菌株.方法 利用物化及基因工程等方法對擬多形假絲酵母菌株的染色體進行改造,將ura3基因和leu2基因進行定嚮誘變,以穫得尿嘧啶和亮氨痠雙營養缺陷型突變菌株,併對其進行生物學鑒定,包括菌株的遺傳穩定性、基因序列及生理生化特徵分析等.結果 篩選得到尿嘧啶和亮氨痠雙營養缺陷型突變菌株,鑒定結果顯示該菌株遺傳穩定性良好,標誌基因均髮生定嚮改變,生理生化結果顯示該菌株能利用多種碳源和氮源,且生長狀況良好.結論 擬多形假絲酵母的雙營養缺陷型工程化改造為進一步研究該菌株的遺傳學奠定瞭基礎,為多價蛋白組成類病毒顆粒的外源錶達提供瞭工程化宿主.進而,為複雜疫苗抗原的重組錶達和功能研究提供瞭技術手段.
목적 이의다형가사효모균(ATCC26012)위재료,구건유전은정성량호적뇨밀정화량안산쌍영양결함형공정화균주.방법 이용물화급기인공정등방법대의다형가사효모균주적염색체진행개조,장ura3기인화leu2기인진행정향유변,이획득뇨밀정화량안산쌍영양결함형돌변균주,병대기진행생물학감정,포괄균주적유전은정성、기인서렬급생리생화특정분석등.결과 사선득도뇨밀정화량안산쌍영양결함형돌변균주,감정결과현시해균주유전은정성량호,표지기인균발생정향개변,생리생화결과현시해균주능이용다충탄원화담원,차생장상황량호.결론 의다형가사효모적쌍영양결함형공정화개조위진일보연구해균주적유전학전정료기출,위다개단백조성류병독과립적외원표체제공료공정화숙주.진이,위복잡역묘항원적중조표체화공능연구제공료기술수단.
Objective To construct a genetically-stable double auxotrophic,in which the uracil and leucine were mutated,using the Candida parapolymorpha ATCC26012 as materials.Methods Based on the physical and genetic engineering methods,the chromosome of the C.parapolymorpha strain was modified,where the ura3 and leu2 genes were directly mutated,to obtain the uracil and leucine double auxotrophic strain.Then the constructed strain was identified by the analysis of its biological properties,such as genetic stability,the change of the genes,and the physiologic and biochemical characteristics.Results The uracil and leucine double auxotrophic strain is obtained by screening.The biological identification results show that the obtained strain is genetically stable and the targeted genes are directly altered.In addition,the physiologic and biochemical analyses indicate that the auxotrophic can utilize various kinds of carbon and nitrogen nutrient sources,and its growth is good.Conclusion The successful construction of double auxotrophic mutant strains facilitated the genetic studies on C.parapolymorpha to meet various investigational purposes.Moreover,the constructed auxotrophic strains can be applied as advantageous host cells to express multiple proteins/antigens simultaneously,which is of great significance in the development of vaccines.
