Bovine pancreatic deoxyribonuclease I produced recombinantly in yeast, Pichia pastoris, to decrease levels of contaminating RNase and eliminate potential pathogens associated with animal based materials.

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Bovine pancreas is a rich source of RNase A which is often found in many commercial DNase preparations. Producing DNase I by recombinant means in an organism with much lower levels of endogenous RNase greatly facilitates purification of an enzyme with undetectable levels of RNase. The processes involved in the production and isolation of recombinant DNase I are completely devoid of animal based components which eliminates the possibility of introducing animal derived pathogens into bioprocessing procedures.

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Recombinant DNase I is suitable for such applications as:

??Removing genomic DNA from RNA preparations prior to RT-PCR

??Degradation of DNA templates after transcription reactions

??Removing unwanted DNA from samples prior to Northern blotting

??Removing DNA during biopharma and bioprocessing procedures

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Unit Definition: One Unit causes an increase in absorbance at 260nm of 0.001 per minute at 25oC when acting upon highly polymerized DNA at pH 5.0, which is the same as other Worthington DNase I products.

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Note: Kunitz units as reported by other suppliers can be 2 to 4 times higher than Kunitz units as measured at Worthington. As measured at Worthington, One Kunitz unit digests 1mg of calf thymus (or pUC19 or l-phage) DNA in 10 minutes at 37oC in 50mM Tris, 1mM Mg2+, 1mM Ca2+, pH 7.8. Correlation of digestion units with Kunitz units may be different in other buffer systems.

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Storage Buffer (DR1S): 5mM calcium acetate, 4mg/ml glycine, pH 5.0 and 50% glycerol.

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DNase I Reaction Buffer (10X): 500mM Tris-HCl, 10mM MgSO4, 1mM CaCl2, pH 7.8, provided.

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艾美捷Worthington脫氧核糖核酸酶I特異性：

bpDNase I?不是堿基或序列特異性的；但是，它不會隨機切割。它顯示優(yōu)先在嘧啶的?5'?側進行裂解，并且在替代共聚物中尤其明顯（Bernardi?等人?1975?和?Lomonossoff?等人?1981）。已經(jīng)表明，扭曲角的變化被?DNase I?識別（Dickerson?和?Drew 1981）。DNase I?的特異性還取決于存在的二價陽離子。在存在?Ca2+?和?Mg2+?的情況下，它會導致單鏈斷裂，而在存在?Mn2+?的情況下會導致雙鏈斷裂（Junowicz?和?Spencer 1973，以及?Campbell?和?Jackson 1980）。

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脫氧核糖核酸酶I相關研究：

肌動蛋白

白蛋白，無核酸酶

脫氧核糖核酸酶?II

脫氧核糖核酸及相關產(chǎn)品

組蛋白

溶菌酶

核酸酶，微球菌

核酸酶，S1

磷酸酶，堿性

磷酸二酯酶?I

磷酸二酯酶?II

蛋白酶K

逆轉(zhuǎn)錄酶，重組?HIV

核糖核酸酶A

核糖核酸酶?T1

核糖核酸酶，E-Rase? RNase?混合物

核糖核酸