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Abstract

Ô½¼òµ¥Ô½ºÃ£¬×ÖÊý¿ØÖÆÔÚ 150¡«250 words.

¾ßÌåÔõÃ´Ð´ÄØ£¿Ïȼòµ¥ÃèÊöһϿÎÌâµÄÇ°ÑØ±³¾°£¬È»ºóÒý³ö×Ô¼ºµÄʵÑ飨 In this paper¡­. or Here, we¡­£©Ö÷Òª³ÂÊöʵÑé½á¹û£¬¿ÉÉÔ´øÌáÒ»ÏÂËùÓõ½µÄÖØÒª·½·¨µÄÃû³Æ£¬È»ºó˵Ã÷ÄãµÄ½á¹ûµÄÒâÒå These data suggest¡­.£»×îºó×ܽá°Î¸ß In a word¡­. or In summary¡­..¡£

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[Background]Acetolactate synthase (ALS) is the key enzyme in isobutanol biosynthetic pathway. Efficient expression of ALS is of great significance for the regulation of isobutanol metabolic pathway. [Methods]The acetolactate synthase gene (alsS) from Bacillus subtilis was amplified by PCR with primers designed according to the sequence of alsS in GeneBank, which is 1 713 bp. Then the alsS was cloned into the expression vector of pET-30a(+). The resulted recombinant plasmid was transformed into Escherichia coli BL2l(DE3) for the overexpression of alsS. [Results]The heterologous expression conditions was optimized to be inducted at an OD600 of 0.6−0.8, 30 ¡ãC with 1 mmol/L IPTG for 6 h . ALS was mostly expressed in the supernatant with the activity of 24.4 U/mL, which was improved for 7.13 times. Electrophoretically pure ALS was obtained after HisTrapTMFF affinity chromatography with the specific activity of 95.2 U/mg. [Conclusion]These results contributed to the construction of isobutanol biosynthetic pathway in E. coli.

Introduction

ÃèÊöÄãËùÑо¿¿ÎÌâµÄÑо¿ÏÖ×´¼°´æÔÚµÄÎÊÌ⣬×ÔÈ»µØ¹ý¶Èµ½ÄãµÄʵÑéÒâÒåºÍÖØÒªÐÔ¡£¡¸Although¡­..there are still¡­ so far¡¹¡¸Here, for the first time, we have used¡­..¡¹¡£Èç¹ûÄãÇ°ÃæÒÑ·¢¹ýÏà¹ØµÄÎÄÕ£¬Òª²åÈëÒýÓÃһϡ¸We recently showed that¡­..¡¹»ò¡¸Our previous studies have shown that¡­ . These results verify that¡­..¡¹¡¸Here¡­..¡¹¡¸ To explore this hypothesis, we generated¡­.¡¹.

дºÃ Introduction Ï൱ÓÚÊÇд×ÛÊö£¬ÐèÒªÔĶÁ´óÁ¿µÄÎÄÏ×½øÐÐÊáÀí×ܽᡣÕâ¹À¼Æ²»ÊÇÌ׸öÄ£°å¾ÍÄÜÁËÊ£¬ÎÒÃÇÏÂÆÚÔÙÁÄ¡£

Materials and Methods

Õⲿ·Ö±È½Ï¼òµ¥£¬»ù±¾ÉÏÊé±¾É϶¼ÓУ¬±ÈÈç PCR¡¢Ã¸»î²â¶¨µÈ£¬½áºÏ×Ô¼ºµÄ¿ÎÌâ×öЩµ÷Õû¼´¿É£¬»òÔòÖ±½Ó˵¾ßÌå¼ûijÎÄÕ»òijÊé¡£

Results

ÕâÒ»²¿·Ö±È½ÏÄÑд¡£¼ÈÒª°´ÕÕÒ»¶¨µÄÂß¼­Ë³Ðò×éÖ¯ÄÚÈÝ£¬ÓÖÒª¶ÔʵÑéÏÖÏó×ö³ö½âÊÍ£¬»¹ÒªÃèÊö׼ȷ¡¢×¨Òµ¡£

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ÃèÊöʵÑé²Ù×÷³£ÓõĶÌÓï¼°¾äʽÓÐ:

In order to prove¡­., we used¡­
To demonstrate¡­., we further¡­
Thus, at current time, we have evidence that¡­
We found that¡­
We have noticed that¡­
It¡¯s known that¡­.
So we introduced¡­.in our study
We next characterized¡­
Next, we examined the effect of¡­
We next set out to determine whether¡­.
Lastly, we examined¡­...

ÃèÊö½áÂÛÐÔÄÚÈÝ»ò³Ð½Ó¹ØÏµ³£ÓõĴÊÓУºNotably ¡¢Importantly¡¢Furthermore¡¢Moreover¡¢Consistently/Consistent with¡¢Compared with¡¢In contrast¡¢As shown in Fig. 1¡¢Fig. 1 shows¡­.¡¢Taken together, these results suggest¡¢These data suggest that¡¢These data are consistent with the notion that.

Discussion

ÕâÒ»²¿·Ö×îÖØÒª£¬Ò²×îÄÑд¡£ÌÖÂÛ²¿·ÖÒ²×îÌåÏÖÒ»¸öÈËд×÷ʵÁ¦£¬Ð´ºÃ·ÇÒ»ÈÕÖ®¹¦£¬¶àÑжÁ´óÅ£ÔÓÖ¾ÄܰïÄãÕÒµ½Ò»Ð©¾äʽºÍ׫д˼·¡£

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¿ªÍ·²¿·Ö£ºOverall, our studies establish the¡­.
Although there are important discoveries revealed by these studies, there are also limitations.

Öмä¹ý³Ì£ºFirst,¡­..Second,¡­..Third,¡­..Fourth,¡­Last,¡­Overall,¡­
Furthermore, our results suggest that
Our studies serve as a proof-of-concept that¡­.This could explain why¡­..Alternatively, ¡­. may be operative¡­.Thus, future iterations of ¡­.. may in fact demonstrate even greater potency

×îºó½áÂÛ£ºn summary, we have identified¡­../Our results confirm that¡­../These studies thus offer a new strategy to treat¡­..

Acknowledgements

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