A Homokaryotic Derivative of a Phanerochaete chrysosporium Strain and Its Use in Genomic Analysis of Repetitive Elements
A Homokaryotic Derivative of a Phanerochaete chrysosporium Strain and Its Use in Genomic Analysis of Repetitive Elements
复制标题
黄孢原平革菌菌株的同核衍生物及其在重复元件基因组分析中的应用
DOI：
--
复制
发表时间：
2000
影响因子：
4.4
通讯作者：
D. Cullen
中科院分区：
文献类型：
--
作者：
P. Stewart;J. Gaskell;D. Cullen
木质素降解菌黄孢原毛革菌的复杂基因家族分析一直受到双核条件的限制。为了便于对黄孢原毛平革菌菌株BKM-F-1767进行遗传学研究，我们从再生的原生质体中分离了一个同核体。通过PCR扩增五个未连锁的基因，然后用等位基因特异性寡核苷酸探测，建立核条件。在标准的氮限制培养条件下，木质素过氧化物酶，锰过氧化物酶，乙二醛氧化酶的活性的同源核体是相当于那些父母的双核体。我们使用同核体来确定基因组组织和评估一个家族的重复元件的转录效应。以前的研究已经确定了一个插入突变，Pce 1，在木质素过氧化物酶等位基因lipI 2。该元件类似于非自主的II类转座子，并且在菌株BKM-F-1767中以多个拷贝存在。在本研究中，另外三个拷贝的Pce 1样元件进行克隆和测序。元素的分布是非随机的，都定位于同一个3.7 Mb的染色体，通过分离分析和Southern印迹分析的同源核。逆转录-PCR（RT-PCR）显示，无论是同核还是亲本双核中，Pce 1都没有从helipI 2转录本中拼接出来。然而，这两个菌株有相当的木质素过氧化物酶活性，这表明一些唇基因可能是多余的。
ABSTRACT Analysis of complex gene families in the lignin-degrading basidiomycete Phanerochaete chrysosporium has been hampered by the dikaryotic nuclear condition. To facilitate genetic investigations in P. chrysosporium strain BKM-F-1767, we isolated a homokaryon from regenerated protoplasts. The nuclear condition was established by PCR amplification of five unlinked genes followed by probing with allele-specific oligonucleotides. Under standard nitrogen-limited culture conditions, lignin peroxidase, manganese peroxidase, and glyoxal oxidase activities of the homokaryon were equivalent to those of the parental dikaryon. We used the homokaryon to determine the genomic organization and to assess transcriptional effects of a family of repetitive elements. Previous studies had identified an insertional mutation, Pce1, within lignin peroxidase allele lipI2. The element resembled nonautonomous class II transposons and was present in multiple copies in strain BKM-F-1767. In the present study, three additional copies of the Pce1-like element were cloned and sequenced. The distribution of elements was nonrandom; all localized to the same 3.7-Mb chromosome, as assessed by segregation analysis and Southern blot analysis of the homokaryon. Reverse transcription-PCR (RT-PCR) showed that Pce1 was not spliced from thelipI2 transcript in either the homokaryon or the parental dikaryon. However, both strains had equivalent lignin peroxidase activity, suggesting that some lip genes may be redundant.
DOI：
10.1073/pnas.86.16.6260
发表时间：
1989
影响因子：
11.1
作者：
Brody,H;Carbon,J
通讯作者：
Carbon,J
真菌中的染色体长度多态性。
DOI：
10.1128/mr.59.4.686-698.1995
发表时间：
1995
期刊：
Microbiological reviews
影响因子：
--
作者：
Zolan,ME
通讯作者：
Zolan,ME
DOI：
10.1073/pnas.94.15.7704
发表时间：
1997-07-22
影响因子：
11.1
作者：
Kidwell, MG;Lisch, D
通讯作者：
Lisch, D