徐州医学院学报
徐州醫學院學報
서주의학원학보
ACTA ACADEMIAE MEDICINAE XUZHOU
2014年
5期
302-305
,共4页
赵金金%徐志辉%胡磊%罗文雅%胡威%尤红娟%李向阳%颜超%郑葵阳%汤仁仙
趙金金%徐誌輝%鬍磊%囉文雅%鬍威%尤紅娟%李嚮暘%顏超%鄭葵暘%湯仁仙
조금금%서지휘%호뢰%라문아%호위%우홍연%리향양%안초%정규양%탕인선
TRAIL%原核表达%纯化%优化
TRAIL%原覈錶達%純化%優化
TRAIL%원핵표체%순화%우화
TNF-related apoptosis inducing ligand%prokaryotic expression%purification%optimization
目的：优化人可溶性TRAIL蛋白生产菌发酵及纯化条件，从而提高TRAIL的产量及活性。方法①探索6种不同培养液、培养液pH值、异丙基硫代半乳糖苷（ IPTG）诱导浓度及时间，选择最佳发酵条件；②联合镍离子亲和层析及超滤法纯化人可溶性TRAIL蛋白。结果①人可溶性TRAIL的最佳发酵条件为pH7．4的YPD培养液，所用IPTG的最佳浓度为0．2 mmol/L，最佳诱导时间为6 h；②经镍离子亲和层析和超滤联合作用可得纯度极高的蛋白。结论成功完成了人可溶性TRAIL发酵及纯化条件的优化，为今后大量提取TRAIL蛋白奠定了基础。
目的：優化人可溶性TRAIL蛋白生產菌髮酵及純化條件，從而提高TRAIL的產量及活性。方法①探索6種不同培養液、培養液pH值、異丙基硫代半乳糖苷（ IPTG）誘導濃度及時間，選擇最佳髮酵條件；②聯閤鎳離子親和層析及超濾法純化人可溶性TRAIL蛋白。結果①人可溶性TRAIL的最佳髮酵條件為pH7．4的YPD培養液，所用IPTG的最佳濃度為0．2 mmol/L，最佳誘導時間為6 h；②經鎳離子親和層析和超濾聯閤作用可得純度極高的蛋白。結論成功完成瞭人可溶性TRAIL髮酵及純化條件的優化，為今後大量提取TRAIL蛋白奠定瞭基礎。
목적：우화인가용성TRAIL단백생산균발효급순화조건，종이제고TRAIL적산량급활성。방법①탐색6충불동배양액、배양액pH치、이병기류대반유당감（ IPTG）유도농도급시간，선택최가발효조건；②연합얼리자친화층석급초려법순화인가용성TRAIL단백。결과①인가용성TRAIL적최가발효조건위pH7．4적YPD배양액，소용IPTG적최가농도위0．2 mmol/L，최가유도시간위6 h；②경얼리자친화층석화초려연합작용가득순도겁고적단백。결론성공완성료인가용성TRAIL발효급순화조건적우화，위금후대량제취TRAIL단백전정료기출。
Objective To optimize the production and purification conditions of human soluble TNF -related apop-tosis inducing ligand ( TRAIL) , thereby increasing the productivity and activity of TRAIL .Methods ①6 different me-dium, pH, the induced concentration and time of isopropyl β-D-1-thiogalactopyranoside ( IPTG) were investigated and the optimum fermentation conditions were selected .②Better human soluble TRAIL protein was obtained through the combination of nickel ion affinity chromatography and ultrafiltration .Results ①The optimum fermentation conditions for human soluble TRAIL was YPD medium at pH 7.4.The optimum concentration of IPTG used was 0.2 mmol/L, the best time was 6 hours.②Through nickel ion affinity chromatography and ultrafiltration combination , higher effects and more purity protein could be obtained .Conclusion The fermentation conditions for human soluble TRAIL are successfully op-timized, which lays foundation for future extraction of large amount of TRAIL protein .
