文中研究了从青霉菌丝体中提取纯化核糖核酸的工艺条件。采用盐碱法破碎细胞,调抽提液 pH值至4.6以沉淀部分蛋白质,然后用中性蛋白酶AS1.398处理分级沉淀后的上清液,详细考察了蛋白酶用量、反应温度、初始pH值和反应时间对核酸纯度的影响。酶解液经截留相对分子质量为6.0×104的超滤膜过滤后,核糖核酸的纯度和得率分别是72.1%和0.42%。
Abstract
The conditions of extraction and purification of RNA from waste Penicillium biomass were studied. A salt and alkali method was used for cell disruption, the pH value of extraction solution was adjusted to 4.6 to precipitate part of
protein, and the supernatant after fractional precipitation was treated with neutral protease AS1.398. The amount of AS1.398 neutral protease, reaction temperature, initial pH value and the hydrolytic time were explored. Then the hydrolysate was treated with ultrafiltration membrane(MWCO is 6.0×104), and the purity and the yield of RNA was 72.1% and 0.42%, respectively.
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1 ] 陈陶声. 氨基酸及核酸类物质发酵生产技术[M] . 北京:化学工业出版社,1993 ,317 - 348
[2 ] 黄明育. 优质核糖核酸生产工艺[J ] . 工业微生物,1993 ,23 (3) :32 - 36
[3 ] Andreu G, Benaiges M D , Lopez J , et al . A simple method for RNA extraction from yeast [J ] . Biotechnol & Bioeng ,1988 ,32(7) :927 - 929
[4 ] 张淑桂,钱敏,王光灿,等. 稀碱法提取酵母RNA 的正交实验研究[J ] . 云南化工,1994 ,10 (2) :10 - 12
[5 ] 廖鲜艳,李永仙,李崎,等. 啤酒酵母胞内RNA 提取的研究[J ] . 酿酒,2002 ,29 (2) :84 - 85
[6 ] 毛宁,洪智勇,张岩芳,等. 利用啤酒酵母提取RNA 的氨法工艺研究Ⅱ[J ] . 药物生物技术,2000 ,7 (1) :35 -37
[7 ] 段作营,于瑞嵩,毛忠贵,等. 谷氨酸菌体中核酸类物质的提取[J ] . 无锡轻工大学学报, 2000 ,19 (4) :322 -324
[8 ] 陈晔. 棉籽水解蛋白和菌体核酸的制备[D] . [学位论文] . 北京:北京化工大学,2003 ,49 - 62
[9 ] 杨健雄. 生物化学与分子生物学实验技术教程[M] .北京:科学出版社,2002 ,35 - 36
[10 ] 杨健雄. 生物化学与分子生物学实验技术教程[M] .北京:科学出版社,2002 ,40 - 42
[11 ] 萨姆布鲁克·J , 弗里奇·E·F , 曼尼阿蒂斯·T. 分子克隆实验指南[M] . 北京:科学出版社,1999 ,304 -316
[12 ] 奥斯伯·F ,金斯顿·R·E ,布伦特·R ,等. 精编分子生物学实验指南[M]. 北京:科学出版社,1999 ,333 - 338
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}