Chinese Journal OF Rice Science ›› 2015, Vol. 29 ›› Issue (6): 571-577.DOI: 10.3969/j.issn.1001G7216.2015.06.002
• Orginal Article • Previous Articles Next Articles
De-zhen WANG, Lu-jia LU, Zhao-jun JIANG, Yu-chun RAO, Mei-hao SUN*()
Received:
2015-02-09
Revised:
2015-07-04
Online:
2015-10-25
Published:
2015-11-10
Contact:
Mei-hao SUN
About author:
*Corresponding author:E-mail:mhsun@zjnu.cn
通讯作者:
孙梅好
作者简介:
*通讯录作者:E-mail:mhsun@zjnu.cn
基金资助:
De-zhen WANG, Lu-jia LU, Zhao-jun JIANG, Yu-chun RAO, Mei-hao SUN. Gene Cloning and Activity Analysis of Adenosine 5'-Phosphosulfate Kinase in Rice[J]. Chinese Journal OF Rice Science, 2015, 29(6): 571-577.
王德珍, 陆璐佳, 姜昭君, 饶玉春, 孙梅好. 水稻腺苷5'-磷酰硫酸激酶编码基因的克隆及其酶活分析[J]. 中国水稻科学, 2015, 29(6): 571-577.
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URL: http://www.ricesci.cn/EN/10.3969/j.issn.1001G7216.2015.06.002
引物 Marker | 引物序列(5'-3') Sequence |
---|---|
OSAPSK1 -F | GAATTCATGGAGCAGCAGCAGC |
OSAPSK1 -R | CTCGAGTTAAGCTTGCAAATAT |
APSK1-C36A-F | CAATATACTGTGGCACAATGCGCCAATTGGACAATCTG |
APSK1-C36A-R | CAGATTGTCCAATTGGCGCATTGTGCCACAGTATATTG |
APSK1-C69A-F | GGGAAAAGCACTCTTGCAGCGGCACTGAATCGGGAG |
APSK1-C69A-R | CTCCCGATTCAGTGCCGCTGCAAGAGTGCTTTTCCC |
Table 1 Primer sequences for OsAPSK1 cloning and mutation.
引物 Marker | 引物序列(5'-3') Sequence |
---|---|
OSAPSK1 -F | GAATTCATGGAGCAGCAGCAGC |
OSAPSK1 -R | CTCGAGTTAAGCTTGCAAATAT |
APSK1-C36A-F | CAATATACTGTGGCACAATGCGCCAATTGGACAATCTG |
APSK1-C36A-R | CAGATTGTCCAATTGGCGCATTGTGCCACAGTATATTG |
APSK1-C69A-F | GGGAAAAGCACTCTTGCAGCGGCACTGAATCGGGAG |
APSK1-C69A-R | CTCCCGATTCAGTGCCGCTGCAAGAGTGCTTTTCCC |
参数 Parameter | KmAPS /(μmol·L-1) | KmATP /(mmol·L-1) | KiATP /(mmol·L-1) | KiAPS /(μmol·L-1) | Kcat /(×103min-1) | Kcat/Km /(×103min-1·μmol-1·L) |
---|---|---|---|---|---|---|
WT | 0.72±0.17 | 0.75±0.18 | 0.61±0.13 | 0.58±0.13 | 0.30±0.042 | 0.41 |
C36A/C69A | 0.44±0.04 | 0.14±0.02 | 0.22±0.04 | 0.68±0.15 | 0.13±0.006 | 0.30 |
Table 2 Comparison of kinetics parameters for OsAPSK1 and mutant C36A/C69A.
参数 Parameter | KmAPS /(μmol·L-1) | KmATP /(mmol·L-1) | KiATP /(mmol·L-1) | KiAPS /(μmol·L-1) | Kcat /(×103min-1) | Kcat/Km /(×103min-1·μmol-1·L) |
---|---|---|---|---|---|---|
WT | 0.72±0.17 | 0.75±0.18 | 0.61±0.13 | 0.58±0.13 | 0.30±0.042 | 0.41 |
C36A/C69A | 0.44±0.04 | 0.14±0.02 | 0.22±0.04 | 0.68±0.15 | 0.13±0.006 | 0.30 |
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