The NIT1 promoter allows inducible and reversible silencing of centrin in Chlamydomonas reinhardtii.
نویسندگان
چکیده
An inverted repeat corresponding to parts of the centrin gene of Chlamydomonas reinhardtii was placed downstream of the NIT1 promoter, which is induced by ammonium starvation. After induction, transformants developed centrin deficiency as assayed by immunofluorescence, Western blotting, and Northern blotting. The effect was reversible, demonstrating that the NIT1 promoter allowed controlled RNA interference in Chlamydomonas reinhardtii.
منابع مشابه
FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii.
In Chlamydomonas reinhardtii, the genes required for nitrate assimilation, including the gene encoding nitrate reductase (NIT1), are subject to repression by ammonia. To study the mechanism of ammonia repression, we employed two approaches to search for mutants with defective repression of NIT1 gene expression. (1) PF14, a gene required for flagellar function, was used as a reporter gene for ex...
متن کاملCentrin scaffold in Chlamydomonas reinhardtii revealed by immunoelectron microscopy.
In the flagellate green alga Chlamydomonas reinhardtii the Ca(2+)-binding EF-hand protein centrin is encoded by a single-copy gene. Previous studies have localized the protein to four distinct structures in the flagellar apparatus: the nucleus-basal body connector, the distal connecting fiber, the flagellar transitional region, and the axoneme. To explain the disjunctive distribution of centrin...
متن کاملCloning of flagellar genes in Chlamydomonas reinhardtii by DNA insertional mutagenesis.
Chlamydomonas is a popular genetic model system for studying many cellular processes. In this report, we describe a new approach to isolate Chlamydomonas genes using the cloned nitrate reductase gene (NIT1) as an insertional mutagen. A linearized plasmid containing the NIT1 gene was introduced into nit1 mutant cells by glass-bead transformation. Of 3000 Nit+ transformants examined, 74 showed mo...
متن کاملInvestigation of an Optimized Context for the Expression of GFP as a Reporter Gene in Chlamydomonas Reinhardtii
Background: Chlamydomonas reinhardtii is a novel recombinant eukaryotic expression system with many advantages including fast growth rate, rapid scalability, absence of human pathogens and the ability to fold and assemble complex proteins accurately, however, obstacle relatively low expression level necessitates optimizing foreign gene expression in this system. The Green Fluorescent Protein (G...
متن کاملA putative protein kinase Femu9p contributes to the iron deficiency-inducible expression of FOX1 gene in Chlamydomonas reinhardtii
FOX1 gene encoding a multicopper ferroxidase is vital in iron metabolism in the single-celled green algae Chlamydomonas reinhardtii. Molecular characteristics of this gene have been extensively studied. However, little information is available about its regulatory factors in C. reinhardtii. In this study, we isolated a regulatory mutant of FOX1 gene by using an arylsulfatase gene driven by the ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Eukaryotic cell
دوره 4 11 شماره
صفحات -
تاریخ انتشار 2005