氯苯降解性质粒pL1限制性酶消化和物理图谱

Restriction endonuclease digestion and physical map of chlorobenzene degradation plasmid pL1

大量提取Ｌ１ 菌质粒并用低熔点琼脂糖回收．用５ 种限制性核酸内切酶消化，所得片段数分别是：ＨｉｎｄⅢ４ ，ＢａｎＨⅠ１６ ，ＸｂａⅠ４ ，ＰｓｔⅠ１５，ＳｍａⅠ１２．从各片段的累加测出该质粒的平均大小约为９２５ ｋｂ ，分子量为６００９ ×１０６ｄａｌｔｏｎ ．在单酶消化的基础上，用Ｈｉｎｄ Ⅲ／ＸｂａⅠ、ＨｉｎｄⅢ／ＢａｍＨⅠ、ＸＢａⅠ／ＢａｍＨⅠ双酶消化，以及Ｂａｍ ＨⅠ部分消化，由此组建了Ｈｉｎｄ Ⅲ／ＸｂａⅠ／Ｂａｍ ＨⅠ３

Plasmid from strain L 1 was prepared on a large and reclaimed by agarose, low melting point. When pL1 DNA was digested by 5 restriction endonucleases, 4 fragments were obtained by Hind Ⅲ, 4 fragments by XbaⅠ, 16 fragments by BamHⅠ,15 fragments by Pst Ⅰ,13 fragments by Sma I.The genome average size was estimated to be 92.5 kb and molecular weight was 60.09×10\+6 Dalton. On the basis of single digestion, pL\-1 DNA was digested with Hind Ⅲ/XbaⅠ,Hind Ⅲ/BamH Ⅰ and XbaⅠ/BamH I,and partially digested with BamH I. The physical map for pL\-1 was constructed from these data.