Himanen J P, Hyvärinen T, Olander R M, Runeberg-Nyman K, Sarvas M
Molecular Biology Unit, National Public Health Institute, Helsinki, Finland.
FEMS Microbiol Lett. 1991 Mar 15;63(1):115-20. doi: 10.1016/0378-1097(91)90538-l.
The subunit S1 of pertussis toxin (PT) was purified as the recombinant product BacS1 from the culture supernatant of a Bacillus subtilis strain containing a secretion vector with a DNA fragment coding for the mature subunit S1 inserted downstream of the signal sequence of the alpha-amylase gene. The method of purification was successive ion exchange and adsorption chromatography. BacS1 occurred in two forms (28 and 20 kDa) of which the truncated 20-kDa peptide was the main one in the supernatant. The truncated BacS1 was purified and shown to have the same NH2-terminus as the full-size (28 kDa) BacS1. It was also enzymatically active indicating correct conformation. The truncated BacS1 was also shown to elicit neutralizing and protective antibodies when injected into mice or rabbits.
百日咳毒素（PT）的亚基S1作为重组产物BacS1，从枯草芽孢杆菌菌株的培养上清液中纯化得到。该菌株含有一个分泌载体，在α-淀粉酶基因信号序列下游插入了编码成熟亚基S1的DNA片段。纯化方法是连续进行离子交换和吸附色谱。BacS1以两种形式（28 kDa和20 kDa）存在，其中截短的20 kDa肽是上清液中的主要形式。截短的BacS1被纯化，并显示其NH2末端与全长（28 kDa）BacS1相同。它也具有酶活性，表明构象正确。当注射到小鼠或兔子体内时，截短的BacS1也能引发中和抗体和保护性抗体。
